Quantitative trait loci mapping of polyphenol metabolites from a 'Latham' x 'Glen Moy' red raspberry (Rubus idaeus L) cross.

OBJECTIVE: The objective of this study was to investigate the genetic control of polyphenol accumulation in red raspberry (Rubus idaeus L). METHODS: The levels of total anthocyanins and 37 individual polyphenol metabolites were measured over three years in a raspberry biparental mapping population. Quantitative trait loci (QTLs) for these traits were mapped onto a high-density SNP linkage map. RESULTS: At least one QTL was detected for each trait, with good consistency among the years. On four linkage groups (LG), there were major QTLs affecting several metabolites. On LG1, a QTL had large effects on anthocyanins and flavonols containing a rutinoside or rhamnose group. On LG4, a QTL had large effects on several flavonols and on LG5 and LG6 QTLs had large effects on ellagic acid derivatives. Smaller QTLs were found on LG2 and LG3. CONCLUSION: The identification of robust QTLs for key polyphenols in raspberry provides great potential for marker-assisted breeding for improved levels of potentially health beneficial components.

The impact of blackcurrant juice on attention, mood and brain wave spectral activity in young healthy volunteers.

There is a growing body of evidence from randomized controlled trials which indicates that consumption of berries has a positive effect upon the cognitive function of healthy adults. It has been recommended that studies combining cognitive and physiological measures be undertaken in order to strengthen the evidence base for the putative effects of flavonoid consumption on cognitive outcomes. This pilot study utilized a randomized, double-blind and placebo controlled crossover design to assess the influence of the acute administration of anthocyanin-rich blackcurrant juice, standardized at 500 mg of polyphenols, on mood and attention. Additionally, this trial used electroencephalography (EEG) to assess if any changes in cognitive performance are associated with changes in localized prefrontal cortex neuronal activity in nine healthy young adults. Outcomes from the pilot EEG data highlight an anxiolytic effect of the consumption of a single serve blackcurrant juice, as indexed by a suppression of α spectral power, and an increase in the slow wave δ and θ spectral powers. There was also an indication of greater alertness and lower fatigue, as indexed by an increase in ß power and suppression of α spectral power. Outcomes from the CogTrack™ system indicated a small acute increase in reaction times during the digit vigilance task.

Antioxidant Rich Potato Improves Arterial Stiffness in Healthy Adults.

Arterial stiffness is an emerging risk factor for cardiovascular disease and dietary anthocyanins may be important in mediating vascular tone. The present study investigated the effect of consumption of an anthocyanin-rich potato, Purple Majesty on arterial stiffness measured as pulse wave velocity in 14 healthy male and female adults. Participants consumed 200 g/day of cooked purple potato containing 288 mg anthocyanins, or a white potato containing negligible anthocyanins for 14 days, separated by a 7-day washout period. Non-invasive assessment of vascular tone by pulse wave velocity was determined in addition to systolic and diastolic blood pressure, high-density lipoproteins, low-density lipoproteins, triglycerides, glucose, insulin and C-reactive protein. Pulse wave velocity was significantly reduced (p = 0.001) following Purple Majesty consumption for 14-days. There were no significant changes with any other clinical parameter measured, and no changes following white potato consumption. The findings from this short-term study indicate a potential effect of Purple Majesty consumption on arterial stiffness.

Polyphenols journey through blood-brain barrier towards neuronal protection.

Age-related complications such as neurodegenerative disorders are increasing and remain cureless. The possibility of altering the progression or the development of these multifactorial diseases through diet is an emerging and attractive approach with increasing experimental support. We examined the potential of known bioavailable phenolic sulfates, arising from colonic metabolism of berries, to influence hallmarks of neurodegenerative processes. In silico predictions and in vitro transport studies across blood-brain barrier (BBB) endothelial cells, at circulating concentrations, provided evidence for differential transport, likely related to chemical structure. Moreover, endothelial metabolism of these phenolic sulfates produced a plethora of novel chemical entities with further potential bioactivies. Pre-conditioning with phenolic sulfates improved cellular responses to oxidative, excitotoxicity and inflammatory injuries and this attenuation of neuroinflammation was achieved via modulation of NF-κB pathway. Our results support the hypothesis that these small molecules, derived from dietary (poly)phenols may cross the BBB, reach brain cells, modulate microglia-mediated inflammation and exert neuroprotective effects, with potential for alleviation of neurodegenerative diseases.

Salal (Gaultheria shallon) and aronia (Aronia melanocarpa) fruits from Orkney: Phenolic content, composition and effect of wine-making.

The polyphenol content and composition of salal and aronia fruits from plants established in Orkney was examined. The composition of the salal fruits has not previously been recorded, and they contained anthocyanins, flavonols, hydroxycinnamates and proanthocyanins. The aronia fruits contained anthocyanins, hydroxycinnamates and flavonols as previously described. Although salal fruits had half the anthocyanin content of aronia fruits, salal wine had higher anthocyanin content, probably due to the relative stability of diglycoside pentose anthocyanins. The wines contained components suggestive of anthocyanin and flavonol degradation, but there was no consistent pattern to stability within phenolic sub-classes. Indeed, the wine made from equal amounts of salal and aronia fruits had patterns of recovery of individual phenolic components which could not be predicted from recoveries in wines from single fruits. This strongly suggests that stability of individual phenolic constituents during wine-making is influenced by the presence and relative stability of other components.

In Vitro Models for Studying Secondary Plant Metabolite Digestion and Bioaccessibility.

There is an increased interest in secondary plant metabolites, such as polyphenols and carotenoids, due to their proposed health benefits. Much attention has focused on their bioavailability, a prerequisite for further physiological functions. As human studies are time consuming, costly, and restricted by ethical concerns, in vitro models for investigating the effects of digestion on these compounds have been developed and employed to predict their release from the food matrix, bioaccessibility, and assess changes in their profiles prior to absorption. Most typically, models simulate digestion in the oral cavity, the stomach, the small intestine, and, occasionally, the large intestine. A plethora of models have been reported, the choice mostly driven by the type of phytochemical studied, whether the purpose is screening or studying under close physiological conditions, and the availability of the model systems. Unfortunately, the diversity of model conditions has hampered the ability to compare results across different studies. For example, there is substantial variability in the time of digestion, concentrations of salts, enzymes, and bile acids used, pH, the inclusion of various digestion stages; and whether chosen conditions are static (with fixed concentrations of enzymes, bile salts, digesta, and so on) or dynamic (varying concentrations of these constituents). This review presents an overview of models that have been employed to study the digestion of both lipophilic and hydrophilic phytochemicals, comparing digestive conditions in vitro and in vivo and, finally, suggests a set of parameters for static models that resemble physiological conditions.

Assessing the influence of genotype and temperature on polyphenol composition in cloudberry (Rubus chamaemorus L.) using a novel mass spectrometric method.

A high-throughput abbreviated liquid chromatography mass spectrometric (ACMS) method was used to assess the relative influence of genotype and temperature on polyphenol composition in cloudberries. Principal component analysis (PCA) plots of the collated ACMS data showed a separation between crosses based on their female parents (Nyby or Fjellgull). Crosses with Nyby as the female parent had higher relative levels of masses assignable to certain ellagitannin derivatives. Crosses with Fjellgull had higher levels of distinctive masses assignable to quercetin derivatives (including a hydroxy-3-methylglutaroyl hexose derivative not previously identified in cloudberry) and anthocyanin derivatives. There was also a separation between samples grown at lower and higher temperatures, which was driven by m/z signals associated with ellagitannins and notably a major component, sanguiin H-6. Therefore, abbreviated MS techniques can discern genetic and/or environmental influences in polyphenol composition and can quickly assess quality in breeding programmes or in response to environmental changes.

Differential effectiveness of berry polyphenols as anti-giardial agents.

Following previous work on the anti-giardial effect of blueberry polyphenols, a range of polyphenol-rich extracts from berries and other fruits was screened for their ability to kill Giardia duodenalis, an intestinal parasite of humans. Polyphenol-rich extracts were prepared from berries using solid-phase extraction and applied to trophozoites of Giardia duodenalis grown in vitro. All berry extracts caused inhibition at 166 µg gallic acid equivalents (GAE)/ml phenol content but extracts from strawberry, arctic bramble, blackberry and cloudberry were as effective as the currently used drug, metronidazole, causing complete trophozoite mortality in vitro. Cloudberry extracts were found to be the most effective causing effectively complete trophozoite mortality at 66 µg GAE/ml. The polyphenol composition of the more effective berry extracts suggested that the presence of ellagitannins could be an important factor. However, the potency of cloudberry could be related to high ellagitannin content but also to the presence of substantial amounts of unconjugated p-coumaric acid and benzoic acid. These in vitro effects occur at concentrations easily achievable in the gut after berry ingestion and we discuss the likelihood that berry extracts could be effective anti-giardial agents in vivo.

In vitro protective effects of colon-available extract of Camellia sinensis (tea) against hydrogen peroxide and beta-amyloid (Aß((1-42))) induced cytotoxicity in differentiated PC12 cells.

There is mounting evidence that the deposition and aggregation of ß-amyloid peptides (Aß) in the brain play a significant role in the development and pathogenesis of Alzheimer's disease. There is further evidence that free radical species such as hydrogen peroxide (H(2)O(2)) mediate Aß induced toxicity. Previous studies have demonstrated that green tea polyphenols possess neuroprotective properties through their ability to ameliorate oxidative stress induced by free radical species. Green tea polyphenols have also been shown to enhance cognition in various animal models of induced cognitive impairment. Upon ingestion, green tea polyphenols are metabolised and undergo bio-transformation which affects their bioavailability and therefore efficacy. In this study, a green tea extract was subjected to a simulated gastrointestinal digestion and a 'colon-available' extract (CAGTE) prepared and assessed for its potential protective effects against H(2)O(2) and Aß((1-42)) induced cytotoxicity using differentiated PC12 cells (dPC12) as a model for neuronal cells. CAGTE represents green tea phytochemicals potentially available after upper gastrointestinal digestion. CAGTE which was depleted in flavan-3-ols, as shown by LC-MS analysis, protected dPC12 cells at concentration ranges of 0.3-10 µg/ml and 0.03-0.125 µg/ml for H(2)O(2) and Aß((1-42)), induced cytotoxicity, respectively. At high concentrations, CAGTE exhibited direct anti-proliferative effects, in line with the reputed anti-cancer properties of green tea polyphenols. These results demonstrate that potentially bioavailable green tea metabolites are able to ameliorate both H(2)O(2) and Aß((1-42)) induced cytotoxicity.

The effect of blueberry extracts on Giardia duodenalis viability and spontaneous excystation of Cryptosporidium parvum oocysts, in vitro.

The protozoan parasites Giardia duodenalis and Cryptosporidium parvum are common causes of diarrhoea, worldwide. Effective drug treatment is available for G. duodenalis, but with anecdotal evidence of resistance or reduced compliance. There is no effective specific chemotherapeutic intervention for Cryptosporidium. Recently, there has been renewed interest in the antimicrobial properties of berries and their phenolic compounds but little work has been done on their antiparasitic actions. The effect of various preparations of blueberry (Vaccinium myrtillus) extract on G. duodenalis trophozoites and C. parvum oocysts were investigated. Pressed blueberry extract, a polyphenolic-rich blueberry extract, and a commercially produced blueberry drink (Bouvrage) all demonstrated antigiardial activity. The polyphenol-rich blueberry extract reduced trophozoite viability in a dose dependent manner. At 167 microgml(-1), this extract performed as well as all dilutions of pressed blueberry extract and the Bouvrage beverage (9.6+/-2.8% live trophozoites remaining after 24h incubation). The lowest dilution of blueberry extract tested (12.5% v/v) contained >167 microgml(-1) of polyphenolic compounds suggesting that polyphenols are responsible for the reduced survival of G. duodenalis trophozoites. The pressed blueberry extract, Bouvrage beverage and the polyphenolic-rich blueberry extract increased the spontaneous excystation of C. parvum oocysts at 37 degrees C, compared to controls, but only at a dilution of 50% Bouvrage beverage, equivalent to 213 microgml(-1) gallic acid equivalents in the polyphenolic-rich blueberry extract. Above this level, spontaneous excystation is decreased. We conclude that water soluble extracts of blueberries can kill G. duodenalis trophozoites and modify the morphology of G. duodenalis and C. parvum.

Memory improvement program for elderly cancer survivors.

Elderly cancer survivors report difficulty with attention, concentration, and memory. This study tested the effectiveness of the Cognitive Behavioral Model of Everyday Memory, an efficacy-based intervention designed to improve memory performance, memory self-efficacy, and metamemory in older adult cancer survivors and those with other chronic conditions. The study provides interesting new evidence that the responses of a group of elders to training varies depending on their health status.

Cell-wall proteins from Sitka spruce xylem are selectively insolubilised during formation of dehydrogenation polymers of coniferyl alcohol.

Extracts from the lignifying xylem of Sitka spruce that were enriched in cell-wall-associated glycoproteins contained peroxidase and oxidase activity and readily formed lignin-like water-insoluble dehydrogenation polymers (DHPs) from coniferyl alcohol (CA) when supplied with H2O2. During the formation of DHPs, the abundance of a number of polypeptides in the extracts was diminished. However, these polypeptides were also diminished in control reactions that contained H2O2 but lacked CA. Polypeptides could be recovered from the DHPs by heating in SDS-PAGE sample buffer but no insolubilised polypeptides could be recovered from the + H2O2 reactions. Although most of the DHP-bound polypeptides were easily removed by pre-washing the DHPs, two polypeptides at 125 and 52 kDa remained tightly bound to the DHPs. The abundance of the two DHP-bound polypeptides mirrored the diminution of 120 and 46 kDa polypeptides in the extracts. The N-terminal protein sequences of the 125 and 52 kDa DHP-bound polypeptides were essentially identical to the sequences obtained from the 120 and 46 kDa polypeptides from the extracts, which confirmed that the DHP-bound polypeptides were derived from these soluble polypeptides. The 125-kDa DHP-bound polypeptide yielded an N-terminal protein sequence that was identical to a laccase-type oxidase previously identified in similar extracts from lignifying Sitka xylem. The N-terminal protein sequence of the 46-kDa polypeptide was homologous with a subset of plant peroxidases. The DHPs had tightly bound peroxidase and oxidase activity, which suggested that these polypeptides were active in their insolubilised state. The mechanism and selectivity of insolubilisation of these enzymes is discussed.

The controversy of prostate screening.

Within the context of aging and ageism, this article evaluates current research on the benefits and risks of prostate cancer screening. Prostate cancer is a concern for many men because it often strikes in the prime of life. Prostate cancer is the most common form of cancer in American men and the second leading cause of cancer-related mortality after lung cancer. Although the disease can be cured if discovered early, it is a slow-growing malignancy that leads to an agonizing death if left untreated. Differential diagnosis often is complicated by comorbid conditions that are part of the normal aging process. Moreover, practitioners' judgment often is prejudiced by common aging misconceptions. Despite the benefits of early screening and detection, the benefits of screening initiatives remain controversial.

Exploring the efficacy of support groups for men with prostrate cancer.

For the 180,000 men who will be diagnosed with prostate cancer this year, improvements in the disease's medical management have prolonged life expectancy. Long-standing treatment side effects include bowel and urinary incontinence and erectile dysfunction, which frequently are embarrassing and contribute to poor emotional well-being. Despite the popularity of support groups in alleviating emotional symptoms related to cancer and its treatment, few men participate in such groups. This article focuses on the reasons why men do not find support groups appealing and presents alternatives that may be more acceptable. Suggestions are made for clinical and research implications.

A comparison of proteins from the developing xylem of compression and non-compression wood of branches of sitka spruce (Picea sitchensis) reveals a differentially expressed laccase.

Soluble and cell wall-associated proteins were extracted from the developing xylem of the compression and non-compression sides of branches of Sitka spruce (Picea sitchensis (Bong) Carr.) by an identical procedure. Equal amounts of proteins were separated by SDS-PAGE, and polypeptides were identified that were more abundant in soluble and cell wall-associated extracts from the developing xylem of either compression or non-compression wood. Two polypeptides (at apparent M(r)s of 48 kDa and 120 kDa) that were more abundant in cell wall-associated extracts of the developing xylem of the compression tissues were selected for amino-terminal protein sequencing. The 48 kDa polypeptide yielded an amino-terminal sequence that had no homology with known protein, gene or EST database sequences. The amino-terminal sequence of the 120 kDa polypeptide was homologous to a number of laccase-type polyphenol oxidases (EC 1.10.3.2) thought to be involved in lignin biosynthesis in trees. Using non-denaturing SDS-PAGE, the 120 kDa laccase was confirmed as a major oxidase activity in extracts of lignifying compression xylem but it was barely detectable in the non-compression extracts where an 85 kDa oxidase was the predominant activity. The differential expression of oxidases in compression and non-compression xylem is discussed.

Oxidase activity in lignifying xylem of a taxonomically diverse range of trees: identification of a conifer laccase.

In a diverse taxonomic range of tree species, including representative species of ancient families of angiosperms (Magnolia x soulangiana Soul.-Bod.) and gymnosperms (Ginkgo biloba L.), oxidase activity was associated with cell walls of developing xylem and was enriched in extracts of cell wall-associated glycoproteins. In all species where oxidase activity was detected histochemically, it was expressed in cell walls of lignifying, differentiating xylem cells and was absent from old wood, cambium and phloem, suggesting that oxidases have a conservative role in lignification of tree xylem. An oxidase from the developing xylem of Picea sitchensis (Bong) Carr. (Sitka spruce) was partially purified by a combination of lectin affinity and immobilized metal ion affinity chromatography. A portion of the total oxidase activity had high affinity for immobilized zinc ions and this feature allowed it to be separated from the bulk of oxidase activity. Two polypeptides that could have been responsible for the bound oxidase activity were enriched by this procedure. The smaller polypeptide of Mr approximately 73 kDa yielded an N-terminal amino-acid sequence that was homologous to laccase-like polyphenol oxidases (E.C. 1.10.3.2) from loblolly pine (Pinus taeda L.), poplar (Populus euramericana (Dode) Guinier) and Arabidopsis. The larger polypeptide (Mr approximately 77 kDa) yielded an N-terminal amino-acid sequence that was homologous with a range of plant subtilisin-like serine proteinases. The roles of oxidase and proteinase activities in developing xylem are discussed.

Fourier-transform infrared and Raman spectroscopic evidence for the incorporation of cinnamaldehydes into the lignin of transgenic tobacco (Nicotiana tabacum L.) plants with reduced expression of cinnamyl alcohol dehydrogenase.

Xylem from stems of genetically manipulated tobacco plants which had had cinnamyl alcohol dehydrogenase (CAD; EC 1.1.1.195) activity down-regulated to a greater or lesser degree (clones 37 and 49, respectively) by the insertion of antisense CAD cDNA had similar, or slightly higher, lignin contents than xylem from wild-type plants. Fourier-transform infrared (FT-IR) microspectroscopy indicated that down-regulation of CAD had resulted in the incorporation of moieties with conjugated carbonyl groups into lignin and that the overall extent of cross-linking, particularly of guaiacyl (4-hydroxy-3-methoxyphenyl) rings, in the lignin had altered. The FT-Raman spectra of manipulated xylem exhibited maxima consistent with the presence of elevated levels of aldehydic groups conjugated to a carbon-carbon double bond and a guaiacyl ring. These maxima were particularly intense in the spectra of xylem from clone 37, the xylem of which exhibits a uniform red coloration, and their absolute frequencies matched those of coniferaldehyde. Furthermore, xylem from clone 37 was found to have a higher content of carbonyl groups than that of clone 49 or the wild-type (clone 37: clone 49: wild-type; 2.4:1.6:1.0) as measured by a degradative chemical method. This is the first report of the combined use of FT-IR and FT-Raman spectroscopies to study lignin structure in situ. These analyses provide strong evidence for the incorporation of cinnamaldehyde groups into the lignin of transgenic plants with down-regulated CAD expression. In addition, these non-destructive analyses also suggest that the plants transformed with antisense CAD, in particular clone 37, may contain lignin that is less condensed (cross-linked) than that of the wild-type.