Blueberry intervention mitigates detrimental microbial metabolite trimethylamine N-oxide by modulating gut microbes.

Gut microbes play a pivotal role in host physiology by producing beneficial or detrimental metabolites. Gut bacteria metabolize dietary choline and L-carnitine to trimethylamine (TMA) which is then converted to trimethylamine-N-oxide (TMAO). An elevated circulating TMAO is associated with diabetes, obesity, cardiovascular disease, and cancer in humans. In the present study, we investigated the effect of dietary blueberries and strawberries at a nutritional dosage on TMA/TMAO production and the possible role of gut microbes. Blueberry cohort mice received a control (C) or freeze-dried blueberry supplemented (CB) diet for 12 weeks and subgroups received an antibiotics cocktail (CA and CBA). Strawberry cohort mice received a control (N) or strawberry-supplemented (NS) diet and subgroups received antibiotics (NA and NSA). Metabolic parameters, choline, TMA, and TMAO were assessed in addition to microbial profiling and characterization of berry powders. Blueberry supplementation (equivalent to 1.5 human servings) reduced circulating TMAO in CB versus C mice (~48%) without changing choline or TMA. This effect was not mediated through alterations in metabolic parameters. Dietary strawberries did not reduce choline, TMA, or TMAO. Depleting gut microbes with antibiotics in these cohorts drastically reduced TMA and TMAO to not-quantified levels. Further, dietary blueberries increased the abundance of bacterial taxa that are negatively associated with circulating TMA/TMAO suggesting the role of gut microbes. Our phenolic profiling indicates that this effect could be due to chlorogenic acid and increased phenolic contents in blueberries. Our study provides evidence for considering dietary blueberries to reduce TMAO and prevent TMAO-induced complications.

Phytochemical profiles, antioxidant activities, and synergistic antiproliferative effects of blueberry and apple peel extracts.

BACKGROUND: Blueberries and apples exhibit favorable bioactivity and health benefits as a result of their rich phytochemicals. Natural phytochemicals exist in complex forms, but there are few reports on whether have additive, synergistic or antagonistic effects between different phytochemicals. The present study aimed to elucidate the synergistic effects of blueberry extract (BE) and apple peel extract (APE) together with respect to inhibiting the proliferation of HepG2 liver cancer cells. Meanwhile, phytochemical characterization of BE and APE was conducted by HPLC, and total antioxidant activity was determined via a cellular antioxidant activity assay, oxygen radical absorption capacity assay and peroxy radical scavenging capacity assay. RESULTS: The results showed that BE and APE were rich in phytochemicals and had potent antioxidant activities, which synergistically inhibited cell proliferation. In the bilateral combination, the dose reduction index value increased by two-fold, and the combination index value at 95% inhibition was less than 1. Additionally, BE + APE supplementation could promote the expression levels of p53 and c-myc genes. In conclusion, the BE and APE had strong antioxidant activity and exhibited synergistic inhibition against proliferation of HepG2 cells. CONCLUSION: The present study can provide a theoretical basis for the synergistic effect of different phytochemicals in health care. © 2023 Society of Chemical Industry.

Inhibition of cell wall pectin metabolism by plasma activated water (PAW) to maintain firmness and quality of postharvest blueberry.

Blueberry is a class of berries with high nutritional and economic value but has short shelf life due to its rapid softening at room temperature. This study investigated the effects of plasma-activated water (PAW) treatment on the softening quality and cell wall pectin metabolism of blueberries stored for 10 d at 25 °C after being immersed in PAW for 10 min. PAW was generated by plasma with different times (1 and 2 min), fixed frequency (10 kHz) and fixed voltage (50 kV). The analysis showed that the firmness of PAW-treated fruit significantly increased (P < 0.05) by 36.4% after 10 d storage. PAW treatment controlled the solubilization of pectin from water-insoluble to water-soluble. The activities of cell wall pectin-degrading enzymes like polygalacturonase (PG), ß-galactosidase (ß-Gal) and pectin methylesterase (PME) in PAW-treated blueberries decreased by 15.7%, 18.3%, and 27.9%, respectively, on day 10. After PAW treatment, blueberries also maintained better postharvest quality (firmness, colour, soluble solid content and anthocyanin content) and intact epidermal waxy and cell wall structure. These results suggested that PAW showed great potential for postharvest fresh-keeping of blueberry.

Strawberry, Blueberry, and Strawberry-Blueberry Blend Beverages Prevent Hepatic Steatosis in Obese Rats by Modulating Key Genes Involved in Lipid Metabolism.

There is an increasing interest in developing natural herb-infused functional beverages with health benefits; therefore, in this study, we aimed to evaluate the effect of strawberry, blueberry, and strawberry-blueberry blend decoction-based functional beverages on obesity-related metabolic alterations in high-fat and high-fructose diet-fed rats. The administration of the three berry-based beverages for eighteen weeks prevented the development of hypertriglyceridemia in obese rats (1.29-1.78-fold) and hepatic triglyceride accumulation (1.38-1.61-fold), preventing the development of hepatic steatosis. Furthermore, all beverages significantly down-regulated Fasn hepatic expression, whereas the strawberry beverage showed the greatest down-regulation of Acaca, involved in fatty acid de novo synthesis. Moreover, the strawberry beverage showed the most significant up-regulation of hepatic Cpt1 and Acadm (fatty acid ß-oxidation). In contrast, the blueberry beverage showed the most significant down-regulation of hepatic Fatp5 and Cd36 (fatty acid intracellular transport). Nevertheless, no beneficial effect was observed on biometric measurements, adipose tissue composition, and insulin resistance. On the other hand, several urolithins and their derivatives, and other urinary polyphenol metabolites were identified after the strawberry-based beverages supplementation. In contrast, enterolactone was found significantly increase after the intake of blueberry-based beverages. These results demonstrate that functional beverages elaborated with berry fruits prevent diet-induced hypertriglyceridemia and hepatic steatosis by modulating critical genes involved in fatty acid hepatic metabolism.

Effects of nitrogen, phosphorus and potassium formula fertilization on the yield and berry quality of blueberry.

Through the application ratio of nitrogen (N), phosphorus (P) and potassium (K) in the field, L9 (33) orthogonal experimental design was used to study the effects of different N, P and K ratios on the yield and quality of blueberry fruit, aiming to optimize the amount of supplied fertilizers. The results showed that N, P and K fertilizer had different effects on fruit yield and quality, among which K fertilizer was the most important factor. Fertilization could significantly improve the yield and fruit quality of blueberry, and the average yield of fertilization treatment was 37.78% higher than that of the control group (CK). Even the treatment with the worst results F6 (N2P3K1), its single fruit weight, anthocyanins, total phenols, soluble solids and soluble protein content were 1.09, 1.32, 1.23, 1.08 and 1.21 times higher than the control (CK), respectively. Based on the comprehensive evaluation of principal component analysis and multi factor analysis of variance, the best fertilization combination for high-yield and good-quality blueberries was N1P2K2 (F2), that is, the best fertilization effect was that including N 100 g/plant, P2O5 25 g/plant, K2O 25 g/plant, applied in the form of ammonium sulfate (472 g/plant), superphosphate (41 g/plant) and potassium sulfate (40 g/plant), respectively.

Blueberry stem extract and stem active components prevent blue light-emitting diode light-induced retinal photoreceptor cell damage in vitro.

Blue light causes retinal damage that can lead to ocular diseases such as age-related macular degeneration. In this study, we determined the protective effect of blueberry stem extract (BStEx) and active components on blue light-emitting diode (LED) light-induced retinal photoreceptor cell damage in vitro. Photoreceptor cells cultured in the presence of BStEx or components were exposed to blue light to induce cell damage. BStEx, fractions of BStEx containing proanthocyanidins, chlorogenic acid, catechin, and epicatechin prevented the cell damage and/or inhibited the generation of reactive oxygen species (ROS). Furthermore, BStEx reduced apoptosis and cell death, and inhibited the phosphorylation of p38 mitogen-activated protein kinase and c-Jun N-terminal kinase leading to cellular apoptosis induced by blue light exposure. These findings suggest that BStEx and components exert a protective effect against blue light-induced photoreceptor cell damage through the inhibition of MAPK phosphorylation and ROS production.

Analysis of Blueberry Plant Rhizosphere Bacterial Diversity and Selection of Plant Growth Promoting Rhizobacteria.

Microbial metabolites in rhizosphere soil are important to plant growth. In this study, microbial diversity in blueberry plant rhizosphere soil was characterized using high-throughput amplicon sequencing technology. There were 11 bacterial phyla and three fungal phyla dominating in the soil. In addition, inorganic-phosphate-solubilizing bacteria (iPSB) in the rhizosphere soil were isolated and evaluated by molybdenum-antimony anti-coloration method. Their silicate solubilizing, auxin production, and nitrogen fixation capabilities were also determined. Eighteen iPSB in the rhizosphere soil strains were isolated and identified as Buttiauxella, Paraburkholderia and Pseudomonas. The higher phosphorus-solubilizing capacity and auxin production in blueberry rhizosphere belonged to genus Buttiauxella sp. The strains belong to genus Paraburkholderia had the same function of dissolving both phosphorus and producing auxin, as well as silicate and nitrogen fixation. The blueberry seeds incubated with the strains had higher germination rates. The results of this study could be helpful in developing the plant growth-promoting rhizobacteria (PGPR) method for enhancing soil nutrients to blueberry plant.

Beneficial effects of blueberry supplementation on the components of metabolic syndrome: a systematic review and meta-analysis.

Metabolic syndrome (MetS) is a combination of interconnected disorders that puts a heavy burden on society. This study investigated the impact of blueberry (BB) supplementation on components of MetS. A systematic search for studies in Embase, Science Direct, Cochrane and PubMed was done. Interventions for at least 2 weeks and studies which investigated the effects of BB on components of MetS in human subjects were included. 25 studies were eligible for inclusion in the review. 21 studies were included in the meta-analysis and the remaining 4 studies in the systematic review. The time range of the assessed studies was from 2007 to 2021. The results of the meta-analysis demonstrated that BB had no significant effect on waist circumference, body mass index (BMI), glycated hemoglobin (HbA1C), glucose level and homeostatic model assessment for insulin resistance (HOMA-IR); however, studies showed a significant improvement in systolic and diastolic blood pressure, triglycerides, total cholesterol, low-density lipoproteins (LDL), high-density lipoproteins (HDL) and insulin levels. In conclusion, the data in this meta-analysis show that BB supplementation is a beneficial option for the management of MetS in humans.

Effect of blueberry extract on energetic metabolism, levels of brain-derived neurotrophic factor, and Ca2+-ATPase activity in the hippocampus and cerebral cortex of rats submitted to ketamine-induced mania-like behavior.

Bipolar disorder (BD) is a psychiatric disease characterized by mood episodes. Blueberry is rich in bioactive compounds and shows excellent therapeutic potential against chronic diseases. The aim of this study was to evaluate the effects of blueberry extract on behavior, energetic metabolism, Ca2+-ATPase activity, and levels of brain-derived neurotrophic factor (BDNF) in the cerebral cortex and hippocampus of rats submitted to an animal model of mania induced by ketamine. Vehicle, lithium (45 mg/kg, twice a day), or blueberry extract (200 mg/kg), was orally administered to Wistar rats for 14 days. Ketamine (25 mg/kg) or vehicle was administered intraperitoneally, once a day, between the 8th and 14th day. On the 15th day, animals received ketamine or vehicle and were subjected to the open field test. Our results demonstrated that the administration of lithium and blueberry extract prevented ketamine-induced hyperlocomotion (P < 0.01). Blueberry extract attenuated the ketamine-induced reduction in the activity of complex I in the cerebral cortex (P < 0.05). Additionally, the administration of ketamine reduced the activities of complexes I and IV (P < 0.05) and citrate synthase in the hippocampus (P < 0.01). However, blueberry extract attenuated the inhibition in the activity of complex IV (P < 0.01). Furthermore, ketamine reduced the Ca2+-ATPase activity in the cerebral cortex and hippocampus (P < 0.05); however, blueberry extract prevented the change in the cerebral cortex (P < 0.05). There were no significant alterations in the levels of BDNF (P > 0.05). In conclusion, this suggested that the blueberry extract can serve as a potential therapeutic strategy for studies searching for novel therapeutic alternatives for BD patients.

Protective Effects of Anthocyanins Extracted from Vaccinium Uliginosum on 661W Cells Against Microwave-Induced Retinal Damage.

OBJECTIVE: To study the protective effect of anthocyanins extracted from Vaccinium Uliginosum (VU) on retinal 661W cells against microwave radiation induced retinal injury. METHODS: 661W cells were divided into 6 groups, including control, model [661W cells radiated by microwave (30 mW/cm2, 1 h)] and VU groups [661W cells pretreated with anthocyanins extracted from VU (25, 50, 100 and 200 µg/mL, respectively) for 48 h, and radiated by microwave 30 mW/cm2, 1 h]. After treatment with different interventions, the cell apoptosis index (AI) was determined using Heochst staining; contents of malonaldehyde (MDA), glutataione (GSH), and activity of superoxide dismutase (SOD) were measured. mRNA expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase 1(HO-1) were detected by real time quantitative polymerase chain reaction, and the expression of HO-1 protein was examined by Western blot analysis. Nucleus and cytoplasm were separated and Nrf2 protein expression was further verified by Western blot analysis. RESULTS: There was significant difference in AI among the groups (F=322.83, P<;0.05). Compared with the control group, AI was significantly higher in the model group and was lower in 4 VU-pretreated groups (P<;0.05). Linear regression analysis showed the decline of AI was in a dose-dependent manner with VU treatment (r=0.8419, P<;0.05). The MDA and GSH contents of 661W cells in VU-treated groups were significantly lower than the model group (P<;0.05). Compared with the model group, the SOD activity in the VU-treated groups (50, 100 and 200 µg/mL) was significantly higher (all P<;0.05). The Nrf2 and HO-1 mRNA expressions were slightly increased after irradiation, and obviously increased in 100 µg/mL VU-treated group. After irradiation, the relative expressions of HO-1 and Nrf2 proteins in nucleus were slightly increased (P<;0.05), and the changes in cytoplasm were not obvious, whereas it was significantly increased in both nucleus and cytoplasm in the VU treatment groups. CONCLUSIONS: Anthocyanins extracted from VU could reduce apoptosis, stabilize cell membrane, and alleviate oxidant injury of mouse retinal photoreceptor 661W cells. The mechanism might be through activating Nrf2/HO-1 signal pathway and inducing HO-1 transcription and translation.

Protective Effects of Anthocyanins Extracted from Vaccinium Uliginosum on 661W Cells Against Microwave-Induced Retinal Damage / 中国结合医学杂志

OBJECTIVE@#To study the protective effect of anthocyanins extracted from Vaccinium Uliginosum (VU) on retinal 661W cells against microwave radiation induced retinal injury.@*METHODS@#661W cells were divided into 6 groups, including control, model [661W cells radiated by microwave (30 mW/cm2, 1 h)] and VU groups [661W cells pretreated with anthocyanins extracted from VU (25, 50, 100 and 200 µg/mL, respectively) for 48 h, and radiated by microwave 30 mW/cm2, 1 h]. After treatment with different interventions, the cell apoptosis index (AI) was determined using Heochst staining; contents of malonaldehyde (MDA), glutataione (GSH), and activity of superoxide dismutase (SOD) were measured. mRNA expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase 1(HO-1) were detected by real time quantitative polymerase chain reaction, and the expression of HO-1 protein was examined by Western blot analysis. Nucleus and cytoplasm were separated and Nrf2 protein expression was further verified by Western blot analysis.@*RESULTS@#There was significant difference in AI among the groups (F=322.83, P<;0.05). Compared with the control group, AI was significantly higher in the model group and was lower in 4 VU-pretreated groups (P<;0.05). Linear regression analysis showed the decline of AI was in a dose-dependent manner with VU treatment (r=0.8419, P<;0.05). The MDA and GSH contents of 661W cells in VU-treated groups were significantly lower than the model group (P<;0.05). Compared with the model group, the SOD activity in the VU-treated groups (50, 100 and 200 µg/mL) was significantly higher (all P<;0.05). The Nrf2 and HO-1 mRNA expressions were slightly increased after irradiation, and obviously increased in 100 µg/mL VU-treated group. After irradiation, the relative expressions of HO-1 and Nrf2 proteins in nucleus were slightly increased (P<;0.05), and the changes in cytoplasm were not obvious, whereas it was significantly increased in both nucleus and cytoplasm in the VU treatment groups.@*CONCLUSIONS@#Anthocyanins extracted from VU could reduce apoptosis, stabilize cell membrane, and alleviate oxidant injury of mouse retinal photoreceptor 661W cells. The mechanism might be through activating Nrf2/HO-1 signal pathway and inducing HO-1 transcription and translation.

Effects of α-casein and ß-casein on the stability, antioxidant activity and bioaccessibility of blueberry anthocyanins with an in vitro simulated digestion.

Blueberry anthocyanins are well-known for their diverse biological functions. However, the instability during digestion results in their weak bioavailability. The current study aimed to investigate the alteration in the stability, antioxidant capacity and bioaccessibility of blueberry anthocyanins with the addition of α-casein and ß-casein in a simulated digestion system using pH differential method, HPLC-MS analysis, peroxyl scavenging capacity (PSC) assay, cellular antioxidant activity (CAA) and penetration test. The results showed that both α-casein and ß-casein could increase the stability of blueberry anthocyanins during intestinal digestion and protect their antioxidant capacity. Moreover, the addition of α-casein or ß-casein would enhance the bioaccessibility of blueberry anthocyanins. In conclusion, our study highlights that the interaction between α-casein or ß-casein with blueberry anthocyanins can protect the compounds against influences associated with the simulated digestion.

[Predicting the Pharmacologic Activity of Natural Materials Based on Metabolomics].

Natural materials such as crude drugs and foods are mixtures composed of various metabolites. Metabolic profiling is often used to identify possible correlations between a compound's metabolic profile and pharmacologic activity. Direct-injection electron ionization-mass spectrometry (DI-EI-MS) is a novel metabolomics method useful for characterizing biological materials. This review demonstrates the establishment of a DI-EI-MS method for metabolic profiling using several closely related lichen species: Cladonia krempelhuberi, C. gracilis, C. pseudogymnopoda, and C. ramulosa. The qualitative DI-EI-MS method was used to profile major and/or minor constituents in extracts of lichen samples. Each lichen sample could be distinguished by altering the DI-EI-MS electron energy and examining the resulting data using one-way analysis of variance. We also attempted to predict pharmacologic activity using DI-EI-MS metabolomics. Blueberry leaf extracts inhibited the proliferation of adult T-cell leukemia (ATL) cells. Blueberry leaf extracts could be distinguished by principal component analysis based on the absolute intensity of characteristic fragment ions. Twenty cultivars were categorized into four species, and the most appropriate discriminative marker m/z value for identifying each cultivar was selected statistically. Components extracted based on DI-EI-MS analyses could be used to construct a model to predict ATL cell bioactivity. These data suggest that the novel DI-EI-MS metabolomics method is suitable for identifying species of natural materials and predicting their pharmacologic activity. This approach could enhance public health by facilitating evaluations of pharmacologic activity and functionality, leading to the elimination of counterfeit products.

Patterns of pollen and resource limitation of fruit production in Vaccinium uliginosum and V. vitis-idaea in Interior Alaska.

Many recent studies assessing fruit productivity of plants in the boreal forest focus on interannual variability across a forested region, rather than on environmental variability within the forest. Frequency and severity of wildfires in the boreal forest affect soil moisture, canopy, and community structure at the landscape level, all of which may influence overall fruit production at a site directly or indirectly. We evaluated how fruit production in two boreal shrubs, Vaccinium uliginosum (blueberry) and V. vitis-idaea (lingonberry), was explained by factors associated with resource availability (such as canopy cover and soil conditions) and pollen limitation (such as floral resources for pollinators and pollen deposition) across boreal forest sites of Interior Alaska in 2017. We classified our study sites into upland and lowland sites, which differed in elevation, soil moisture, and active layer. We found that resource and pollen limitation differed between the two species and between uplands and lowlands. Lingonberry was more pollen limited than blueberry, and plants in lowland sites were more pollen limited relative to other sites while plants in upland sites were relatively more resource limited. Additionally, canopy cover had a significant negative effect in upland sites on a ramet's investment in reproductive tissues and leaves versus structural growth, but little effect in lowland sites. These results point to importance of including pollinator service as well as resource availability in predictions for changes in berry abundance.

Effect of atmospheric cold plasma treatment on antioxidant activities and reactive oxygen species production in postharvest blueberries during storage.

BACKGROUND: Blueberry is universally acknowledged as a kind of berry rich in antioxidants. Cold plasma, an emerging non-thermal treatment technology, has been proved to be able to maintain or improve the antioxidant level while inactivating the microorganisms on the surface of fruits and vegetables. Postharvest blueberries were treated with atmospheric cold plasma (ACP; 12 kV, 5 kHz) for 0 s (Control), 30 s (ACP-30), 60 s (ACP-60), and 90 s (ACP-90) in this study, and the effects of ACP on the antimicrobial properties, antioxidant activities, and reactive oxygen species (ROS) production were investigated during storage at 4 ± 1 °C for 40 days. RESULTS: Total aerobic bacteria and mold populations on ACP-treated blueberries decreased significantly in a time-dependent manner (P < 0.05), and decreased by 0.34-1.24 and 0.57-0.87 log10 CFU g-1 respectively on ACP-60-treated blueberries during storage. The decay rate of blueberries was decreased by 5.8-11.7% and the decrease of blueberry firmness was slowed down by ACP-60. But the total phenol, anthocyanin, and ascorbic acid contents increased, and superoxide dismutase, catalase, and peroxidase activities were enhanced in ACP-treated blueberries. The free radical scavenging activity and total antioxidant capacity (T-AOC) were enhanced. Hydrogen peroxide (H2 O2 ) and superoxide anion (O2 - ) production rates declined by 27.3% and 41.3% at day 40 of storage, respectively. CONCLUSION: It is suggested that ACP may be a promising non-thermal treatment technology for postharvest sterilization and preservation of blueberry under suitable conditions. © 2020 Society of Chemical Industry.

Valorization of Persimmon and Blueberry Byproducts to Obtain Functional Powders: In Vitro Digestion and Fermentation by Gut Microbiota.

Globalization of fruit and vegetable markets generates overproduction, surpluses, and potentially valuable residues. The valorization of these byproducts constitutes a challenge, to ensure sustainability and reintroduce them into the food chain. This work focuses on blueberry and persimmon residues, rich in polyphenols and carotenoids, to obtain powders with high added value to be used as ingredients in food formulation. These powders have been characterized, and the changes in the bioactive compounds in in vitro gastrointestinal digestion have been evaluated. The results indicated that the type of residue, the drying process, as well as the content and type of fiber determine the release of antioxidants during digestion. In vitro colonic fermentations were also performed, and it was observed that the characteristics of digested powders had an effect on the composition of the growing microbial community. Thus, carotenoids and anthocyanins maintain an interplay with microbiota that could be beneficial for human health.

Ethylene plays an important role in the softening and sucrose metabolism of blueberries postharvest.

We studied the effects of ethylene on softening and sucrose metabolism in postharvest blueberry fruit by examining the responses of fruit firmness, cell wall polysaccharides, cell wall enzymes, four key genes of cell wall degradation and metabolism, enzyme activities, and five key genes of sucrose metabolism to exogenous ethylene treatments. Ethylene was found to accelerate blueberry softening, as it promoted the degradation of pectin and expression of pectinesterase (PE) and polygalacturonase (PG). Sucrose catabolism was accelerated with fruit softening, while sucrose content, sucrose phosphate synthase (SPS) activity were positively correlated with the loss of fruit firmness. Exogenous ethylene treatments promoted sucrose metabolism by inhibiting the expression of VcSPS1 and VcNIN2 and stimulating the expression of VcSS1 and VcCWINV1. These results indicate that ethylene plays an important role in fruit softening and sucrose metabolism of blueberry at 20 °C, and there may be a link between sucrose metabolism and fruit softening.

Combinatorial effect of blueberry extracts and oxaliplatin in human colon cancer cells.

The use of natural compounds to potentiate the effect of drugs and lower their adverse effects is an active area of research. The objective is to determine the effect of combined blueberry extracts (BE) and oxaliplatin (OX) in colon cancer cells. The results demonstrated that treatments of BE/OX showed inhibitory effects on HCT-116 cell and nontoxic effect on CCD-18Co normal colon cells. Flow cytometry analysis indicated that treatment with the BE, OX or in combination could induce G0/G1 cell cycle arrest, apoptosis, increase of reactive oxygen species, and induce loss of mitochondrial membrane potential in HCT-116 cells. Furthermore, after treatments, the expression of inflammatory cytokines was decreased, cyclin D1 and CDK4 were decreased; caspases-3 and 9 were activated; the Akt/Bad/Bcl-2 pathway was modulated. Moreover, the combination treatment had a considerably higher growth inhibitory effect on human colon cancer HCT-116 cells than that of BE or oxaliplatin alone. Our results showed that BE increased the anticolon cancer effect of OX making it an attractive strategy as adjuvant therapy to potentially reduce the adverse side effects associated with chemotherapeutic drugs.

Direct-Injection Electron Ionization-Mass Spectrometry Metabolomics Method for Analyzing Blueberry Leaf Metabolites That Inhibit Adult T-cell Leukemia Proliferation.

Metabolic profiling is often used to identify possible correlations between a compound's metabolic profile and biological activity. Direct-injection electron ionization-mass spectrometry "fingerprinting" is useful for characterizing biological materials. We demonstrate the utility of direct-injection electron ionization-mass spectrometry for metabolic profiling using 100 different extracts of leaves from 20 blueberry cultivars collected at 5 time points from April to December 2008. A qualitative direct-injection electron ionization-mass spectrometry method was used to profile the major and/or minor constituents in the blueberry leaf extracts. Blueberry leaf extracts could be distinguished by principal component analysis based on the absolute intensity of characteristic fragment ions. Twenty cultivars were categorized into four species, and the most appropriate discriminative marker m/z value for identifying each cultivar was selected statistically. Correlated m/z values indicating the collection month were determined in the same analysis, and air temperature variance factors were extracted from score plots by principal component analysis. We previously reported that blueberry extracts inhibit the proliferation of adult T-cell leukemia cells. Leaves of Vaccinium virgatum collected in December of 2008 exhibited significantly greater inhibition of adult T-cell leukemia cell proliferation than other species. Highly bioactive cultivars or species were identified by direct-injection electron ionization-mass spectrometry metabolomics analysis of blueberry leaf extracts. The components extracted based on our direct-injection electron ionization-mass spectrometry analyses could be used to construct a model to predict anti-adult T-cell leukemia bioactivity. This is the first study to report a relationship between seasonal variation and bioactivity of natural products using a direct-injection electron ionization-mass spectrometry metabolomics method.

Transcriptional Activation of Anthocyanin Biosynthesis in Developing Fruit of Blueberries ( Vaccinium corymbosum L.) by Preharvest and Postharvest UV Irradiation.

The effect and mechanism of preharvest and postharvest ultraviolet (UV) irradiation on anthocyanin biosynthesis during blueberry development were investigated. The results showed that preharvest UV-B,C and postharvest UV-A,B,C irradiation significantly promoted anthocyanin biosynthesis and the transcripts of late biosynthetic genes (LBG) VcDFR, VcANS, VcUFGT, and VcMYB transcription factor as well as DFR and UFGT activities in anthocyanin pathway in a UV wavelength- and developmental stage-dependent manner. VcMYB expression was positively correlated with that of VcANS and VcUFGT and coincided with anthocyanin biosynthesis responding to the UV radiation. Sugar decreased during postharvest but increased during preharvest UV radiation in mature fruit. Our results indicate that UV-responsive production of anthocyanins is mainly caused by the activation of anthocyanin downstream pathway genes, which could be upregulated by VcMYB. Furthermore, different potential response mechanisms may exist between preharvest and postharvest UV radiation in blueberries, involving a systemic response in living plants and a nonsystemic response in postharvest fruit.