Chemical Characterization of Pruning Wood Extracts from Six Japanese Plum (Prunus salicina Lindl.) Cultivars and Their Antitumor Activity.

The Japanese plum tree (Prunus salicina Lindl.) is mainly cultivated in temperate areas of China and some European countries. Certain amounts of wood (from pruning works) are generated every year from this crop of worldwide commercial significance. The main objective of this work was to value this agricultural woody residue, for which the chemical composition of pruning wood extracts from six Japanese plum cultivars was investigated, and the antiproliferative activity of extracts and pure phenolics present in those extracts was measured. For the chemical characterization, total phenolic content and DPPH radical-scavenging assays and HPLC‒DAD/ESI‒MS analyses were performed, with the procyanidin (-)-ent-epicatechin-(2α→O→7,4α→8)-epicatechin (5) and the propelargonidin (+)-epiafzelechin-(2ß→O→7,4ß→8)-epicatechin (7) being the major components of the wood extracts. Some quantitative differences were found among plum cultivars, and the content of proanthocyanidins ranged from 1.50 (cv. 'Fortune') to 4.44 (cv. 'Showtime') mg/g of dry wood. Regarding the antitumoral activity, eight wood extracts and four phenolic compounds were evaluated in MCF-7 cells after 48 h of induction, showing the wood extract from cv. 'Songold' and (‒)-annphenone (3), the best antiproliferative activity (IC50: 424 µg/mL and 405 µg/mL, respectively).

Drying date plum (Diospyros lotus L.) fruit: Assessing rehydration properties, antioxidant activity, and phenolic compounds.

Date plum (Diospyrus lotus L.) is an edible fruit from the Ebenaceae family, rich in nutrients, and having tremendous medicinal properties. This paper attempted to show the influence of different parameters of convective drying such as temperature (50, 60, 70, and 80°C) and air velocity (0.5, 1.0, and 1.5 m/s) on the shrinkage and microstructure, rehydration properties, antioxidant activity, and phenolic compounds of date plum. The drying caused significant changes in the color, actual size, and distribution of the fruit cells of date plum. The total phenolic content (TPC), total flavonoid content (TFC), ferric reducing antioxidant power (FRAP), and 2,2-diphenyl-1-picrylhydrazyl (DPPH) of fresh date plum were 0.81 ± 0.00 mg GAE/g, 0.23 ± 0.10 mg ECE/g, 7.15 ± 1.09 mmol ISE/g, and 14.92 ± 0.88 mmol/TE, respectively. The drying at 70°C had the highest values of TPC, TFC, gallic acid, chlorogenic and syringic acids, catechin, quercetin-3-glucoside, resveratrol, and DPPH. The drying air velocities showed no significant effects on the antioxidant contents and the antioxidant activity. Of the models applied to the drying kinetics, the Midilli model was found as the best model to describe the drying kinetics of date plum. In addition, the Weibull model was found as the most successful among the models applied to the rehydration kinetics of date plum. According to the achieved findings, the convective drying temperature of 70°C is the optimum temperature to produce the dehydrated date plum. Practical Application This work has revealed the drying conditions responsible for preserving the phenolic compounds, total flavonoid content, and antioxidant features of D. lotus L. The study found the optimum drying conditions, and Midilli and Weibull models were the most fitted models to describe the drying and rehydration behaviors of D. lotus L. fruits, respectively. The drying provides a reasonable value of the possibility of continuous consumption of the fruits dried afforded on off-seasons. The dried fruits are widely used for multipurpose and have been extensively used in food industries due to their rich nutraceutical and antioxidant compounds.

Co-Ingestion of Natal Plums (Carissa macrocarpa) and Marula Nuts (Sclerocarya birrea) in a Snack Bar and Its Effect on Phenolic Compounds and Bioactivities.

This study investigated the effect of co-ingesting Natal plums (Carissa macrocarpa) and Marula nuts (Sclerocarya birrea) on the bioaccessibility and uptake of anthocyanins, antioxidant capacity, and the ability to inhibit α-glucosidase. A Natal plum-Marula nut bar was made by mixing the raw nuts and the fruit pulp in a ratio 1:1 (v/v). The cyanidin-3-O-sambubioside (Cy-3-Sa) and cyanidin-3-O-glucoside content (Cy-3-G) were quantified using the ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC/Q-TOF-MS). Inclusion of Natal plum in the Marula nut bar increased the Cy-3-Sa, Cy-3-G content, antioxidants capacity and α-glucosidase inhibition compared to ingesting Marula nut separately at the internal phase. Adding Natal plum to the Marula nut bar increased bioaccessibility of Cy-3-Sa, Cy-3-G, quercetin, coumaric acid, syringic acid and ferulic acid to 80.2% and 71.9%, 98.7%, 95.2%, 51.9% and 89.3%, respectively, compared to ingesting the Natal plum fruit or nut separately.

Polyphenolic QTOF-ESI MS Characterization and the Antioxidant and Cytotoxic Activities of Prunus domestica Commercial Cultivars from Costa Rica.

There is an increased interest in plum research because of their metabolites' potential bioactivities. In this study, the phenolic profiles of Prunus domestica commercial cultivars (Methley, Pisardii and Satsuma) in Costa Rica were determined by Ultra Performance Liquid Chromatography coupled with High Resolution Mass Spectrometry using a quadrupole-time-of-flight analyzer (UPLC-ESI-QTOF MS) on enriched phenolic extracts obtained through Pressurized Liquid Extraction (PLE) under acidic and neutral extraction conditions. In total, 41 different phenolic compounds were identified in the skin and flesh extracts, comprising 11 flavan-3-ols, 14 flavonoids and 16 hydroxycinnamic acids and derivatives. Neutral extractions for the skins and flesh from all of the cultivars yielded a larger number of compounds, and were particularly rich in the number of procyanidin trimers and tetramers when compared to the acid extractions. The total phenolic content (TPC) and antioxidant potential using the DPPH and ORAC methods exhibited better results for neutral extracts with Satsuma skins and Methley flesh, which showed the best values (685.0 and 801.6 mg GAE/g extract; IC50 = 4.85 and 4.39 µg/mL; and 12.55 and 12.22 mmol TE/g extract, respectively). A Two-Way ANOVA for cytotoxicity towards AGS gastric adenocarcinoma and SW620 colon adenocarcinoma indicated a significant difference (p < 0.05) for PLE conditions, with better results for neutral extractions, with Satsuma skin delivering the best results (IC50 = 60.7 and 46.7 µg/mL respectively) along with Methley flesh (IC50 = 76.3 and 60.9 µg/mL, respectively). In addition, a significant positive correlation was found between TPC and ORAC (r = 0.929, p < 0.05), as well as a significant negative correlation (p < 0.05) between TPC and cytotoxicity towards AGS and SW620 cell lines (r = -0.776, and -0.751, respectively). A particularly high, significant, negative correlation (p < 0.05) was found between the number of procyanidins and cytotoxicity against the AGS (r = -0.868) and SW620 (r = -0.855) cell lines. Finally, the PCA clearly corroborated that neutral extracts are a more homogenous group exhibiting higher antioxidant and cytotoxic results regardless of the part or cultivar; therefore, our findings suggest that PLE extracts under neutral conditions would be of interest for further studies on their potential health benefits.

Anthocyanins attenuate vascular and inflammatory responses to a high fat high energy meal challenge in overweight older adults: A cross-over, randomized, double-blind clinical trial.

BACKGROUND & AIMS: Postprandial metabolic imbalances are important indicators of later developing cardiovascular disease (CVD). This study investigated the effects of food anthocyanins on vascular and microvascular function, and CVD associated biomarkers following a high fat high energy (HFHE) meal challenge in overweight older adults. METHODS: Sixteen subjects (13 female, 3 male, mean age 65.9 SD 6.0 and body mass index 30.6 kg/m2 SD 3.9) participated in a crossover, randomized, controlled, double-blind clinical trial (registered under Australian New Zealand Clinical Trials Registry, identifier no. ACTRN12620000437965). Participants consumed a HFHE meal with a 250 mL dose of either intervention (anthocyanins-rich Queen Garnet Plum) or control (apricot) juice. Blood samples and blood pressure measures were collected at baseline, 2 h and 4 h following the HFHE meal. Vascular and microvascular function were evaluated at baseline and 2 h after the HFHE meal. RESULTS: Participants had a higher 2 h postprandial flow-mediated dilatation (+1.14%) and a higher microvascular post-occlusive reactive hyperaemia (+0.10 perfusion units per mmHg) when allocated to the anthocyanin compared to the control arm (P = 0.019 and P = 0.049, respectively). C-reactive protein was lower 4 h postprandially in the anthocyanins (1.80 mg/L, IQR 0.90) vs control arm (2.30 mg/L, IQR 1.95) (P = 0.026), accompanied by a trend for lower concentrations of interleukin-6 (P = 0.075). No significant postprandial differences were observed between treatments for blood pressure, triacylglycerol, total cholesterol, serum derivatives of reactive oxidative metabolites, tumor necrosis factor alpha, interleukin-1 beta, or maximum microvascular perfusion following iontophoresis of acetylcholine. CONCLUSION: Fruit-based anthocyanins attenuated the potential postprandial detrimental effects of a HFHE challenge on parameters of vascular and microvascular function, and inflammatory biomarkers in overweight older adults. Anthocyanins may reduce cardiovascular risk associated with endothelial dysfunction and inflammatory responses to a typical high fat 'Western' meal. Further studies are required to better elucidate the clinical implications of postprandial biomarkers of CVD.

Pectin-based edible coating preserves antioxidative capacity of plum fruit during shelf life.

Exploiting safer methods for fruit preservation such as application of edible coatings can improve shelf life, valuable characteristics, and antioxidative capacity. The current study aimed to investigate the effect of a pectin-based edible coating on antioxidative capacity of plum fruit during shelf life (19 ± 2 ℃ and 65% relative humidity for eight days). To do this, three solutions (0.5, 1, and 1.5%) of pectin, plasticized by glycerol (0.3% w/v), were applied on plum fruit and compared to a control treated with only distilled water. Ascorbic acid, total phenolics, anthocyanin and flavonoid contents, total antioxidative capacity based on 1,1-diphenyl-2-picryl-hydrazyl hydrate method, peroxidase (as an antioxidant enzyme), and polyphenol oxidase (as an oxidant enzyme) activities were recorded during this period. The results demonstrated that pectin-based edible coating was significantly effective on maintaining ascorbic acid, anthocyanin and flavonoid contents, and antioxidative capacity in plum fruits (P ≤ 0.01). The activities of enzymes were significantly affected by the coatings; peroxidase activity increased and polyphenol oxidase activity decreased (P ≤ 0.01). All pectin concentrations significantly caused higher ascorbic acid and anthocyanin contents, antioxidative capacity, and peroxidase activity but a lower polyphenol oxidase activity than the control; however, just 1 and 1.5% concentrations were effective in terms of total phenolic compounds and flavonoid content, respectively, and the other concentrations acted the same as the control. In general, the coating constituted from 1.5% pectin showed the best results for most measured parameters. Considering the influences of pectin-based edible coating on antioxidative characteristics of plum fruits, its application can be potentially regarded as a favorable method to enhance nutritional value of fruits.

Physico-Chemical Properties and Oxidative Stability of Fixed Oil from Plum Seeds (Prunus domestica Linn.).

Storage of a great amount of plum kernel waste becomes a challenge for food industry. In this work, the plum seed was used as a source of fixed oil that can be an ingredient of commercial products. Soxhlet extraction was carried out using the different solvents, such as n-hexane, n-heptane, ethyl acetate, acetone, or chloroform:methanol mixture (2:1 v/v). The highest yield of oil (about 30%) was obtained using n-heptane and n-hexane, while the lowest yield was obtained using ethyl acetate. The analysis of physico-chemical parameters indicated that all samples of plum seed oil have an exceptional quality. Schaal oven test indicated that the fixed oil of plum seed exhibited satisfactory oxidative stability at moderate storage temperatures (up to 65 °C). The composition of phenolic compounds in the oil samples was determined using HPLC method. The most abundant compound of seven identified and quantified phenolic compounds was vanillic acid. The highest content of ß-carotene (1.67 mg 100 g-1 fixed oil) spectrophotometrically determined was in the oil extracted with n-hexane. The lowest content of ß-carotene (1.26 mg 100 g-1 fixed oil) was determined in the oil extracted with a mixture of chloroform:methanol (2:1 v/v). This oil had the highest antioxidant activity (IC50 value of 4.35 mg mL-1) compared to other oil samples. The antioxidant activity was probably caused by the presence of phenolic compounds. The investigated physico-chemical properties demonstrated that the plum seed oil has a potential for application in the food, cosmetics, and pharmaceutical industry.

New acylated cyanidin glycosides extracted from underutilized potential sources: Enzymatic synthesis, antioxidant activity and thermostability.

Interest in anthocyanins has increased remarkably in recent decades, although their wider application has been hampered by instability problems. Thus, this study aimed at developing a strategy to gain access to more stable anthocyanins via enzymatic esterification. For that purpose, three cyanidin derivatives were obtained from underutilized, but easily accessible sources, and their total anthocyanin content was quantified. The purity of cyanidins obtained ranged from 40% to 88% depending on their source. Subsequently, the critical enzymatic reaction conditions were established, and the best results were found using tert-butanol as a solvent, 20 g/L of lipase B from Candida Antarctica, and vinyl cinnamate as acyl donor at ratio 250:1 (acyl donor to anthocyanin). Finally, five new acylated anthocyanin derivatives were synthesized with improved antioxidant activity and thermostability, in comparison to the cyanidin-3-glucoside, which is an advantageous feature for industrial applications.

Qualitative and Quantitative Evaluation of Heat-Induced Changes in Polyphenols and Antioxidant Capacity in Prunus domestica L. By-products.

Plum pomace, an agro-industrial waste product has received attention due to the worldwide popularity of plums. During convection, the content of flavan-3-ols decrease, except drying at 90 °C, whereas the content of i.e., chlorogenic, 3-p- and 4-p-coumaroylquinic acids, quercetin rutinoside, and galactoside was observed to increase along with the increase in process temperature. The highest content of all identified polyphenols was found in plum pomace powders obtained using a combination of convective at 90 °C and microwave vacuum drying (MVD) at 120 W, whereas the highest retention of the group consisted of phenolic acids, flavonols, and anthocyanins was noted when CD 70 °C/MVD 120 W was used, pointing to a strong influence of the type of polyphenols on their changes caused by drying. The correlations between TEAC ABTS and the sum of flavonoids (r = 0.634) and anthocyanins (r = 0.704) were established. The multiple regression analysis showed that polyphenol content was more strongly affected by drying time than by maximum temperature, whereas antioxidant capacity was more influenced by maximum temperature of sample than by drying time.

Polyphenol-Enriched Plum Extract Enhances Myotubule Formation and Anabolism while Attenuating Colon Cancer-induced Cellular Damage in C2C12 Cells.

Preventing muscle wasting in certain chronic diseases including cancer is an ongoing challenge. Studies have shown that polyphenols derived from fruits and vegetables shows promise in reducing muscle loss in cellular and animal models of muscle wasting. We hypothesized that polyphenols derived from plums (Prunus domestica) could have anabolic and anti-catabolic benefits on skeletal muscle. The effects of a polyphenol-enriched plum extract (PE60) were evaluated in vitro on C2C12 and Colon-26 cancer cells. Data were analyzed using a one-way ANOVA and we found that treatment of myocytes with plum extract increased the cell size by ~3-fold (p < 0.05) and stimulated myoblast differentiation by ~2-fold (p < 0.05). Plum extract induced total protein synthesis by ~50% (p < 0.05), reduced serum deprivation-induced total protein degradation by ~30% (p < 0.05), and increased expression of Insulin-Like Growth Factor-1 (IGF-1) by ~2-fold (p < 0.05). Plum extract also reduced tumor necrosis factor α (TNFα)-induced nuclear factor κB (NFκB) activation by 80% (p < 0.05) in A549/NF-κB-luc cells. In addition, plum extract inhibited the growth of Colon-26 cancer cells, and attenuated cytotoxicity in C2C12 myoblasts induced by soluble factors released from Colon-26 cells. In conclusion, our data suggests that plum extract may have pluripotent health benefits on muscle, due to its demonstrated ability to promote myogenesis, stimulate muscle protein synthesis, and inhibit protein degradation. It also appears to protect muscle cell from tumor-induced cytotoxicity.

Nutrient, fibre, sorbitol and chlorogenic acid content of prunes (Prunus domestica): an updated analysis and comparison of different countries of origin and database values.

Current prune composition data are outdated and require a comprehensive and comparative re-analysis. This novel study aimed to: (i) analyse and compare prune composition from major countries of origin; and (ii) provide a comprehensive compositional analysis of prunes of USA origin and compare this with UK and USA database data. Prune samples were analysed for major nutrients and bioactive compounds and compared between countries of origin. Total fibre was higher in prunes from the USA (12.0 g/100 g) and Chile (11.5 g/100 g) compared with France (8.4 g/100 g) and Argentina (8.9 g/100 g), while prunes from all countries contained high levels of sorbitol (11.2-15.5 g/100 g). Differences in energy and starch values compared with national databases reflected different approaches to sampling and analysis. In conclusion, prunes contain high levels of fibre and other bioactive compounds. Variations between country of origin and database values highlight the importance of transparency in documenting sampling and analysis methods.

Evaluation of Polyphenolics Content and Antioxidant Activity in Edible Wild Fruits.

Plant diversity is a basic source of food and medicines for the local communities of the Himalayas. Current study was intended to assess polyphenolics content and antioxidant potential in edible wild fruits used as food and to treat various diseases by the inhabitants of Himalayan region of Pakistan. The fruits of 20 plant species were evaluated using standard protocols, whereas information on medicinal uses was gathered through semistructured interviews. Comparatively, Prunus domestica and Rubus ellipticus fruits exhibited highest levels of phenolics and flavonols contents (113.55 ± 0.61 mg GAE/100 g and 200.06 ± 1.57 mg RtE/100 g FW, respectively) in acetone extract. Nevertheless, flavonoids were maximum in the water extract of Rosa moschata (194.82 ± 3.46 mg RtE/100 g FW). Contrary, Duchesnea indica fruit depicted significant potential to scavenge DPPH and H2O2 radicals at 94.66 ± 8.89% in acetone extract and 83.54± 9.37% in water extract, while acetone extract of Rubus ellipticus had maximum potential to reduce ferric ions (133.66 ± 15.00 µM GAE/100 g FW). Additionally, total antioxidant capacity was highest in the acetone extract of Berberis lycium fruit (332.08 ± 21.90 µM AAE/100 g FW). The relationships between polyphenolics and antioxidant activity revealed synergistic role of secondary metabolites in the prevention of diseases. Our study revealed that wild fruits consumed by the local communities of Himalayas are rich in health beneficial phytochemicals and hold significant potential to treat chronic diseases, particularly associated with free radicals.

Facile synthesis of fluorescent carbon dots from Prunus cerasifera fruits for fluorescent ink, Fe3+ ion detection and cell imaging.

Carbon dots (CDs) synthesized from natural products have drawn numerous attentions due to some unique properties. Here, Prunus cerasifera fruits were used as carbon source to synthesize high luminescent CDs by hydrothermal method. The obtained CDs were characterized by TEM, FTIR and XPS methods, founding the CDs were near-spherical and contained abundant nitrogen element. The CDs aqueous solution exhibited bright blue fluorescence under ultraviolet illumination, with the maximum emission at 450 nm. They could be potentially used as invisible fluorescent ink by written on the paper and irradiated by UV light, due to their fluorescent properties. Moreover, the CDs were found being selectively quenched by Fe3+ ion. The quench of CDs was linearly related to the concentration of Fe3+ ion in the range of 0-0.5 mM, meaning they could be developed as fluorescent probe of Fe3+ ion. At last, the CDs were used for cell imaging, founding they were low toxicity to HepG2 cells and exhibited blue and green fluorescence under a fluorescence microscope. In summary, the CDs prepared from Prunus cerasifera fruits exhibited excellent fluorescence properties, and could be potentially applied in the field of fluorescent ink, Fe3+ ion detection and cell imaging.

Impact of year of harvest, genotype and cultivation method on bioactives and Pru d 1 allergen content in plums.

The present work studied the effect of the year of harvest, the genotype and the cultivation method on the nutritional quality and the allergen content of three plum cultivars. The common quality parameters and the phytochemical content strongly varied with the year and the cultivar, while the system of cultivation had a minor influence. In particular, ascorbic acid greatly decreased in 2016 compared to 2015, while polyphenols were higher in 2016. The health-promoting compounds, and particularly phenolics, were significantly correlated with the antioxidant capacity. Finally, the allergen content was strongly dependent on the content of flavan-3-ols, suggesting that this class of phenolics is determinant in influencing the allergen content in plums. Results showed that the major factor affecting the quality and the concentration of natural metabolites of plum, in addition to the diversity among genotypes, is the year-to-year variation, whereas the system of cultivation plays a marginal role.

Oxalic acid preharvest treatment increases antioxidant systems and improves plum quality at harvest and during postharvest storage.

BACKGROUND: Plums are much appreciated by consumers as fresh fruit but have a limited storage life. Since reactive oxygen species (ROS) are associated with fruit ripening, an increase in ROS scavenging antioxidant systems could lead to a delay in postharvest plum ripening and in maintaining fruit quality after long cold storage. RESULTS: Results showed that crop yield (kg per tree) and fruit weight were enhanced by preharvest oxalic acid (OA) treatment of plum cultivars ('Black Splendor' and 'Royal Rosa'), although the on-tree ripening process was delayed. In addition, the ripening process during cold storage was delayed in plums from OA-treated tress, manifested by lower firmness and acidity losses and reduced ethylene production, as compared with fruits from control trees. Antioxidant compounds (phenolics, anthocyanins and carotenoids) and the activity of antioxidant enzymes were higher in plums from OA-treated trees than in controls, at harvest and during 50 days of cold storage. CONCLUSION: OA preharvest treatment could be a useful tool to maintain plum quality properties during long-term storage, by delaying the postharvest ripening process through a delay in ethylene production, with an additional effect on increasing bioactive compounds with health beneficial effects. © 2018 Society of Chemical Industry.

Antioxidative Capacity of and Contaminant Concentrations in Processed Plum Products Consumed in Romania.

The bioactive compounds (total phenols, anthocyanins, and flavonoids) and antioxidant activity of plum-based products (prunes, jams, and marmalade) were evaluated based on spectrophotometric assays and acrylamide and 5-hydroxymethyl-2-furaldehyde concentrations to determine the quality and safety of these products in the Romanian marketplace. The total polyphenol concentrations in prunes, marmalade, and jams were 0.93 to 5.63 g of gallic acid equivalent per kg of dry matter, and the antioxidant activity was 3.0 to 17.2 mmol/kg. The concentrations of potentially harmful acrylamide in the plum products ranged from the limit of quantitation (<20 µg/kg) to 37.44 µg/kg, and the concentration was higher in prunes (26.66 to 163.72 µg/kg) than in jams. The 5-hydroxymethyl-2-furaldehyde concentrations in plum products were also variable, at 36 to 2,149 mg/kg. These data revealed a very low concentration of acrylamide in thermally processed plum products, and the results for total polyphenols and the antioxidant activity of prunes and plum jams may be relevant for evaluating these products as important sources of bioactive compounds.

Plum (Prunus salicina) peel and pulp microparticles as natural antioxidant additives in breast chicken patties.

Fiber microparticles (MPCs) separately obtained from peel and pulp of Japanese plum residues contained co-extracted ß-carotene, lutein, and α- and γ-tocopherols, as well as polyphenols (cyanidins, quercetin derivatives, pentameric proanthocyanidins). Peel and pulp MPCs were then separately evaluated as natural antioxidant additives (2.0% w/w level) in raw breast chicken patty, susceptible to oxidation. Their effect on technological properties was also analyzed. MPCs reduced in 50% the formation of thiobarbituric acid reactive substances (TBARS) in raw patties during 10-days storage at 4.0°C. Ferric reducing power (FRAP) was 77-157% higher in MPCs-added patties, especially with peel MPCs, being then attributed to the antioxidants supplied by these MPCs. It can be also associated to the highest α- and γ-tocopherol levels found in the peel MPCs-added patties, which remained high after cooking as well. Also, higher pectin and low lignin contents of pulp MPCs determined greater hydration, stabilized the cyanidins and, hence, the red color transferred to raw patties, and increased springiness of cooked patties. Plum peel and pulp MPCs are efficient additives for chicken meat products.

Select polyphenolic fractions from dried plum enhance osteoblast activity through BMP-2 signaling.

Dried plum supplementation has been shown to enhance bone formation while suppressing bone resorption. Evidence from previous studies has demonstrated that these responses can be attributed in part to the fruit's polyphenolic compounds. The purpose of this study was to identify the most bioactive polyphenolic fractions of dried plum with a focus on their osteogenic activity and to investigate their mechanisms of action under normal and inflammatory conditions. Utilizing chromatographic techniques, six fractions of polyphenolic compounds were prepared from a crude extract of dried plum. Initial screening assays revealed that two fractions (DP-FrA and DP-FrB) had the greatest osteogenic potential. Subsequent experiments using primary bone-marrow-derived osteoblast cultures demonstrated these two fractions enhanced extracellular alkaline phosphatase (ALP), an indicator of osteoblast activity, and mineralized nodule formation under normal conditions. Both fractions enhanced bone morphogenetic protein (BMP) signaling, as indicated by increased Bmp2 and Runx2 gene expression and protein levels of phosphorylated Smad1/5. DP-FrB was most effective at up-regulating Tak1 and Smad1, as well as protein levels of phospho-p38. Under inflammatory conditions, TNF-α suppressed ALP and tended to decrease nodule formation (P=.0674). This response coincided with suppressed gene expression of Bmp2 and the up-regulation of Smad6, an inhibitor of BMP signaling. DP-FrA and DP-FrB partially normalized these responses. Our results show that certain fractions of polyphenolic compounds in dried plum up-regulate osteoblast activity by enhancing BMP signaling, and when this pathway is inhibited by TNF-α, the osteogenic response is attenuated.

Dried plum alleviates symptoms of inflammatory arthritis in TNF transgenic mice.

Dried plum (DP), a rich source of polyphenols has been shown to have bone-preserving properties in both animal models of osteoporosis and postmenopausal women. We evaluated if DP alleviated the destruction of joints in transgenic mice (TG) that overexpress human tumor necrosis factor (TNF), a genetic model of rheumatoid arthritis (RA). A four-week treatment of 20% DP diet in TG slowed the onset of arthritis and reduced bone erosions in the joints compared to TG on a regular diet. This was associated with fewer tartrate-resistant acid phosphatase (TRAP) positive cells, suggesting decreased osteoclastogenesis. A DP diet also produced significant protection of articular cartilage and reduction of synovitis. Cultures of human synovial fibroblast in the presence of TNF showed a significant increase in inflammatory interleukin (IL)-1ß, chemokines (monocyte chemoattractant protein-1: MCP1 & macrophage inflammatory protein-1 alpha: MIP1α), cartilage matrix metalloproteinases (MMP1&3), and an osteoclastogenic cytokine (receptor activator of nuclear factor-κB ligand: RANKL) compared to controls. Addition of neochlorogenic acid (NC), a major polyphenol in DP to these cultures resulted in down-regulation of these genes. In the cultures of mouse bone marrow macrophage, NC also repressed TNF-induced formation of osteoclasts and mRNA levels of cathepsin K and MMP9 through inhibition of nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1) expression and nuclear factor kappa B (NF-κB) activation. Our data suggested that dietary supplementation with DP inhibited TNF singling; leading to decreased erosions of bone and articular cartilage as well as synovitis.