Enzymatic synthesis of Vaccinium blue using vaccinoside as a bifunctional precursor.

Since Tang dynasty in China, the fresh leaves of Vaccinium bracteatum (VBL) have been applied as natural pigment to produce black rice. However, detailed information on its biosynthetic mechanism still remained unclear. Following rice dyeing capacity assay, vaccinoside, one of iridoid glycosides, was identified as the key active compound. Increased methodical research demonstrated vaccinoside as a distinct bifunctional precursor, which could be catalyzed by polyphenol oxidase or ß-glucosidase independently, followed by reaction with 15 amino acids to give blue pigments (VBPs; λmax 581-590 nm) of different hues. Two synthetic pathways of VBPs were proposed, using multiple techniques such as HPLC, HPSEC, UV-Vis spectrum and colorimeter as analysis tools. Black rice was interpreted to be prepared by cooking, using vaccinoside, intrinsic enzymes from fresh VBL and rice protein in combination. These findings promote the understanding of VBP formation mechanisms and provide an efficient method of producing novel Vaccinium blue pigments.

HPLC-Mass spectrometry analysis of phenolics comparing traditional bilberry and blueberry liqueurs.

The purpose of this study was to investigate the differences between the phenolics compositions of bilberry and blueberry liqueurs, to determine whether these are comparable. Both bilberry and blueberry liqueurs have the same name on the market, however they differ in phenolic composition and content. A total of 43 compounds were identified, 36 in the bilberry liqueur, and 21 in the blueberry liqueur. High performance liquid chromatography coupled with mass spectrometry (HPLC-MS) was used to identify and quantify these compounds, where they were fragmented to the MS4 fragments. Anthocyanins were the major phenolics group detected in the bilberry liqueurs, but only the second most abundant phenolics group in the blueberry liqueurs. The most abundant phenolics group in the blueberry liqueurs was hydroxycinnamic acids. The lowest content of individual phenolic compounds was for blueberry liqueur made from whole fruit. This study will provide valuable data to consumers in their choice of a more expensive bilberry liqueur or a cheaper blueberry liqueur.

Optimisation of Polyphenols Extraction from Wild Bilberry Leaves-Antimicrobial Properties and Stability Studies.

Polyphenolic extracts from natural sources have received great interest due to their beneficial properties for human health. A method to reduce their variability is to use the design of experiments which allows a limited number of experiments to be performed while exploring the experimental space. Firstly, a 23-full factorial model was used to investigate the polyphenols extraction from wild bilberry leaves. Spectrophotometric data (the content of polyphenols, flavonoids, chlorophyll and radical scavenger activity) and extraction yield were used as responses, and six statistical models were determined depending on the two numerical factors (temperature and alcohol % of ethanol-water mixture) being significant (p < 0.05) in all cases. Numerical optimisation performed by Design Expert 13 software correlates well with the chemical profile determined by high-performance liquid chromatography and the amount of the polyphenol. Afterwards, under the optimised conditions, an extract was prepared in three extraction steps for which composition, chemical stability and antimicrobial properties were evaluated. The antimicrobial potential of the extract was compared with that of the standard compounds (rutin and chlorogenic acid), and the results supported a synergistic effect of the extract components.

Inhibitory Potential of Different Bilberry (Vaccinium myrtillus L.) Extracts on Human Salivary α-Amylase.

Recently, consumer preferences for bilberries have increased markedly. This fact is probably related to their natural constituents, such as phenolic compounds including anthocyanins and tannins, as well as the vitamins and minerals they contain. Phenolic compounds are known for their numerous beneficial effects on human health. Moreover, bilberry fruits have been shown to inhibit the activity of carbohydrate hydrolyzing enzymes, which can significantly decrease the postprandial increase in blood glucose levels. Thus, the aim of the present study is to investigate the inhibitory effect of Vaccinium myrtillus L. extracts on key enzyme α-amylase, linked to type 2 diabetes. No data have been published on the inhibitory properties of Vaccinium myrtillus L. fruits growing wild in Bulgaria against carbohydrate enzymes. Bilberry extracts were analyzed for total polyphenols, total anthocyanin content, antioxidant activity and their inhibitory properties against α-amylase. The contents of flavonols, anthocyanins and stilbenes were determined by HPLC analysis. The identified flavonols in the analyzed bilberry extracts were mainly represented by quercetin derivatives as rutinoside. The predominant anthocyanins for both aqueous and organic solvents were delphinidin-3-galactoside and malvidin-3-glucoside. The results revealed that bilberry extracts are effective inhibitors of α-amylase, with IC50 values from 20.8 to 194.8 µg GAE/mL. All the samples proved to have antioxidant activity measured by three different in vitro assays (FRAP, CUPRAC and DPPH). The inhibitory properties of V. myrtillus L. extracts may provide a new direction in the development and research of new pharmaceuticals for the suppression of postprandial hyperglycemia in diabetic patients.

The Action Mechanisms, Anti-Cancer and Antibiotic-Modulation Potential of Vaccinium myrtillus L. Extract.

BACKGROUND: Herbal medicinal products containing Vaccinium myrtillus L. (bilberry) fruits and fruit extracts are widely available in the market. Although bilberry leaves and stems are considered as bio-waste, they contain much higher levels of phenolic compounds than fruits. The study aimed to investigate the antimicrobial and anticancer potential of aerial part extracts from Vaccinium myrtillus L. (V. myrtillus, VM) plants harvested at high altitudes in Armenian landscape and characterize the bioactive phytochemicals. MATERIAL AND METHODS: For evaluation of antioxidant properties, chemical-based tests (total phenolic and flavonoid content, and antiradical activity in 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) tests) and cellular antioxidant activity (CAA) assay were applied. Genotoxicity and anticancer properties of the extract alone and in combination with fluorouracil were explored in human cancer and normal cell lines. Antibacterial properties of V. myrtillus extract alone and in combination with antibiotics, as well as their effect on proton-flux rate through cell membrane were explored on bacterial strains. The characterization of active phytochemicals was done using Liquid Chromatography-Quadrupole-Orbitrap High-Resolution Mass Spectrometry (LC-Q-Orbitrap HRMS). RESULTS: The V. myrtillus aerial part extract demonstrated promising antioxidant properties in all tests. The selective cytotoxic activity was documented against various cancer cell lines (human colon adenocarcinoma (HT29), human breast cancer (MCF-7) and human cervical carcinoma (HeLa)), while it did not inhibit the growth of tested human normal primary renal mixed epithelial cells (HREC) even at 10-fold higher concentrations. The extract did not have genotoxic properties in comet assay making it a potential source for the development of anticancer preparations. The investigated extract did not directly inhibit the growth of Escherichia coli (E. coli) and Salmonella typhimurium (S. typhimurium) strains at up to 1 mg/mL concentration. However, V. myrtillus extract enhanced the kanamycin intake and increased its efficiency against E. coli strain. The phytochemical characterization of the extract showed the presence of different groups of phenolics. CONCLUSIONS: Based on obtained data, we suggest the aerial parts of the V. myrtillus plant as an alternative source of bioactive natural products for food supplements, nutraceuticals, functional foods and medicine.

Traceability of bilberries (Vaccinium myrtillus L.) of the Baltic-Nordic region using surface-enhanced Raman spectroscopy (SERS): DFT simulation-based DNA analysis.

Food traceability is a major issue in the industry. We investigated whether bilberries (Vaccinium myrtillus L.) from 4 different locations within the Baltic-Nordic region could be effectively differentiated using surface-enhanced Raman scattering (SERS) based spectral data and chemometric analyses. Furthermore, we aimed to determine if nucleobase (adenine and cytosine) methylation could be responsible for any observed variation. Our experiment was successful in that both principal component (PCA) and discriminant function analyses (DFA) showed differentiation between bilberry DNA from all 4 geographical regions. Density functional theory (DFT) based simulations allowed us to analyze whether DNA's spectral data dissimilarities may be due to nucleobase methylation. Although results were inconclusive on this, our investigation provides valuable data on simulated versus experimental DNA and DNA component spectra. Further research will be directed towards understanding what other epigenetic changes could be responsible for the observed DNA variation as well as determining the optimal parameters for using DFT simulations in upcoming projects.

Quality by Design in optimizing the extraction of (poly)phenolic compounds from Vaccinium myrtillus berries.

Quality by Design was adopted for developing an effective extraction procedure of (poly)phenolic compounds from bilberry (Vaccinium myrtillus L.) fruits, using a pooled sample of berries from different regions of Ukraine. Mechanical solvent extraction, microwave-assisted extraction (MAE) and ultrasonic-assisted extraction (UAE) were investigated by screening matrices. Extraction time (Time, from 5 to 15 min), organic solvent type (OS type, methanol, ethanol and acetone), organic solvent percentage (OS%, from 50% to 90%), sample/extractant ratio (S/E ratio, from 0.025 to 0.1 g mL-1), and, only for MAE, extraction temperature (T, from 30 to 60°C), were selected as critical method parameters (CMPs). The spectrophotometric assays total soluble polyphenols (TSP), total monomeric anthocyanins (TMA), and radical scavenging activity (evaluated by the 2,2-diphenyl-1-picrylhydrazyl radical (DPPH), the 2,2'-azino-di-(3-ethylbenzthiazoline sulfonic acid), and the ferric reducing antioxidant power methods) were chosen as critical method attributes (CMAs). The screening procedure allowed for selecting UAE and methanol, while the other CMPs underwent further optimization through Response Surface Methodology. Target values for TSP, TMA and DPPH were selected and the method operable design region (MODR) was defined by means of Monte-Carlo simulations. The optimized conditions, with the corresponding MODR intervals in bracket, were the following: (i) Time, 17 min (15-23 min); OS%, 56% (44-59%); S/E ratio, 0.030 (0.022-0.034) g mL-1. Under these experimental conditions, CMAs values of the pooled sample were the following (n = 3): TSP=4433±176 mg (+)-catechin eq/100 g dry weight (d.w.); TMA=3575±194 mg cyanidin-3-glucoside eq/100 g d.w.; DPPH=273±5 µg DPPH inhib./mg d.w. The optimized extraction method was tested for matrix effect (ME%) in the UHPLC-MS/MS analysis of 15 anthocyanins and 20 non-anthocyanins individual (poly)phenols commonly found in bilberries, as well as for luteolin, sinapic acid, and pelargonidin-3-glucoside, absent in this fruit and therefore added to the extracts as surrogate standards for evaluating apparent recovery (AR%). |ME%| was in any case ≤ 23% and AR% of the surrogate standards in the range 91-95%, confirming the very good performances of the optimized extraction method.

Protective Effect of Bilberry Anthocyanin Extracts on Dextran Sulfate Sodium-Induced Intestinal Damage in Drosophila melanogaster.

Inflammatory bowel disease (IBD) is a chronic recurrent disease that can be controlled by various natural extracts. Anthocyanins (ANCs) from bilberry have significant antioxidant capacity and are widely used as food colors and antioxidants. In this study, we investigated the protective effects of bilberry anthocyanin extracts (BANCs) against dextran sulphate sodium (DSS)-induced intestinal inflammation in a Drosophila melanogaster (D. melanogaster) model, and the effects on the lifespan, antioxidant capacity, intestinal characteristics, and microbiome and gene expression profiles were analyzed to elucidate the underlying biological mechanisms. In DSS-induced normal and axenic D. melanogaster, BANCs significantly increased the survival rate, maintained the intestinal morphology and integrity, and reduced the number of dead intestinal epithelial cells and the ROS level of these cells. BANC supplementation had no significant effect on the intestinal microflora of DSS-induced D. melanogaster, as demonstrated by a 16S rDNA analysis, but improved the antioxidant capacity by activating the relative gene expression of NRF2 signaling pathways in the intestine of D. melanogaster with DSS-induced inflammation. Therefore, the results demonstrate that BANCs effectively alleviate intestinal inflammatory injury induced by DSS and improve the antioxidant capacity of D. melanogaster by modulating NRF2 signaling pathways, and could thus promote the application of BANCs as functional foods.

Influence of the time of bilberry (Vaccinium myrtillus L.) addition on the phenolic and protein profile of beer.

BACKGROUND: In recent years, there has been an increased fruit addition to the brewing process, especially in craft breweries. Fruit addition leads to changes in the organoleptic characteristics and chemical composition of beers. Bilberries are rich in phenolic compounds and possess significant antioxidant capacity. The effects of bilberry addition and brewing process parameters on the changes in the phenolic and protein profile of beer have not been sufficiently studied. The aim of this research was to investigate the changes in the individual phenolic compounds and the protein fractions in beer when bilberries were added at different maturation stages. METHODS: An infusion mashing method was applied for the purpose of obtaining wort with an original extract of 14°P after boiling. Pilsner malt, bitter and aromatic hops 60/40 (Perle and Cascade, respectively), dry yeast Saccharomyces pastorianus (carlsbergensis) Saflager W 34/70, and bilberries (Vaccinium myrtillus L.) were used. All processes were conducted in a Home Brew 50 all-in-one 50 dm3 brewing system. The fermentation was carried out in a stainless steel cylindroconical fermenter at a temperature of 14°C. The “green beer” was transferred to small stainless-steel fermenters after 60% of the original wort extract had been fermented. The maturation continued for 14 days at 14°C, and the beer lagering for 5 days at 2°C. The bilberries were pasteurised in a water bath for 10 minutes at 70°C. After cooling, they were added to small fermenters at a concentration of 167 g/dm3 at the beginning and on the seventh day of beer maturation. All variants were carried out in duplicate. After lagering, the beer was bottled using a “beer gun”. The beer samples from the experiments were filtered on the day of bottling and frozen until analysis. HPLC/UV-VIS and electrophoresis were used to determine the phenolics and proteins, respectively. The total monomeric anthocyanins were determined by the pH differential method. The original wort extract and alcohol concentration were evaluated, and the sensory analysis was performed according to EBC standard methods. RESULTS: The changes in 10 phenolic acids, 7 flavonoids, and 10 protein fractions in beer with bilberries added at the beginning and on the seventh day of maturation were studied. The addition of bilberries led to an increase in the phenolic acid (3-fold) and flavonoid (6.2-fold) concentrations. The highest enrichment was observed in terms of rutin, chlorogenic, caffeic, and 3,4-dihydrobenzoic acids. Rosmarinic acid and monomeric anthocyanins were only detected in the bilberry beers. Chlorogenic and neochlorogenic acids, rutin, and catechin dominated in the bilberry beers. Neochlorogenic and gallic acids, epicatechin, and catechin dominated in the bilberry-free beers. The addition of bilberries reduced the protein content by 93 to 96%. The number of protein fractions decreased from 10 to 4. The influence of the bilberry addition time on the phenolics and proteins was different, and it affected the individual protein fractions in a different way. More phenolic acids and flavonoids were determined when bilberries were added at the beginning of maturation. The losses of some protein fractions were higher at the beginning of maturation and of others on the seventh day of maturation, whereas the addition time did not matter for some of the fractions. CONCLUSIONS: This study provides new information related to the changes in the phenolic and protein profile of beer with bilberries depending on the time of bilberry addition during beer maturation. The protein concentration and number of protein fractions decreased dramatically. In spite of the significant protein losses, the bilberry addition improved the phenolic profile of the beer and its organoleptic characteristics. The presence of more phenolic compounds is related to the antioxidant capacity respective to the biological value of beer. Further research in this direction is needed.

Phytochemical and pharmacological anti-diabetic properties of bilberries (Vaccinium myrtillus), recommendations for future studies.

Nowadays, extensive attention has focused on dietary constituents that may be valuable for treating, eating, and preventing diabetes. Numerous studies have shown that anthocyanin's are one of the most important nutritional factors associated with diabetes. Anthocyanin's are the leading group of water-soluble pigments in the plant kingdom, and they are generally available in some human diet in fruits, vegetables, cereals, beans. Amongst, bilberries (Vaccinium myrtillus), is one of the essential sources for dietary anthocyanin consumption containing vast amounts of anthocyanin's, making them the main plant in the treatment and prevention of diabetes. Although the bilberries have other valuable properties such as anti-cancer, anti-inflammatory, and antioxidant, the main focus of the present study is to present the effects of bilberries (V. myrtillus) on the prevention and treatment of diabetes.

The effect of Vaccinium bracteatum Thunb. leaves addition on antioxidant capacity, physicochemical properties, and in vitro digestibility of rice extrudates.

Vaccinium bracteatum Thunb.leaves (VBTL) are a traditional Chinese herbal medicine with potential natural pigments and antioxidants. Its application information on extrusion is limited. In this study, to increase the usage of foods containing VBTL, rice-based extrudates with VBTL were investigated. The extrusion was performed at a wide temperature range (60-140°C) and with different VBTL supplementations (3%-10%). The extrudates with 10% VBTL showed a significant decrease in total phenolic content (TPC), total flavonoid content (TFC), and antioxidant activity upon extrusion. TPC and TFC increased with the rise of barrel temperature. The VBTL color was stable during the wide-temperature extrusion process. The bioactivities of the extrudates were positively correlated with the VBTLsupplement (R = 0.943-0.989). In addition, different addition levels of VBTL significantly changed the water absorption index (WAI), water solubility index (WSI), pasting properties, and in vitro starch digestibility of the extrudates. Moreover, extrudates with 3% VBTL showed the highest WAI and peak viscosity but the lowest in vitro starch digestibility. These results may suggest that rice extrudates with a low supplement of VBTL have an acceptable color and may be used as functional additives for high antioxidant and low glycemic response diets. PRACTICAL APPLICATION: The results showed that the Vaccinium bracteatum Thunb. leaves (VBTL) color and antioxidation were stable during the wide-temperature extrusion process. Extrusion processing can be used to enlarge the usage of VBTL and simplify its traditional processing method. Applying VBTL to the process of extrusion appears to be a suitable method of producing value-added and low glycemic response extrudates.

Application of Crude Pomace Powder of Chokeberry, Bilberry, and Elderberry as a Coloring Foodstuff.

Berry pomace, rich in polyphenols, especially anthocyanins, accumulates during the production of red juices. Pomace from chokeberry (Aronia melanocarpa Michx.), bilberry (Vaccinium myrtillus L.), and elderberry (Sambucus nigra L.) represent good sources of coloring foodstuffs. Pomace powders (PP) were prepared by milling the seedless fractions of the three dried berry pomaces (50 °C, 8 h). Techno-functional properties of the powders such as particle size distribution, bulk density, sedimentation velocity, and swelling capacity were determined to evaluate the powders for possible food applications. Total anthocyanin content was quantified by UHPLC-DAD before and during a storage experiment to monitor the degradation of anthocyanins in the PP and in a yogurt model application. The high content of phenolic compounds and the still intact cell structure ensured high stability of anthocyanins over 28 days of storage. In the model application, color saturation was stable over the whole storage time of 14 days. Regarding the techno-functional properties, only a few differences between the three PP were observed. The particle size of elderberry PP was larger, resulting in lowest bulk density (0.45 g/mL), high cold-water solubility (16.42%), and a swelling capacity of 10.16 mL/g dw. Sedimentation velocity of the three PP was fast (0.02 mL/min) due to cluster formation of the particles caused by electrostatic and hydrophobic properties. Compared to other high-intensity coloring foodstuffs, the use of PP, showing acceptable color stability with potential health-promoting effects, represents a wide applicability in different food applications and especially in products with a longer shelf-life.

Effects of Totum-63 on glucose homeostasis and postprandial glycemia: a translational study.

Global prevalence of type 2 diabetes (T2D) is rising and may affect 700 million people by 2045. Totum-63 is a polyphenol-rich natural composition developed to reduce the risk of T2D. We first investigated the effects of Totum-63 supplementation in high-fat diet (HFD)-fed mice for up to 16 wk and thereafter assessed its safety and efficacy (2.5 g or 5 g per day) in 14 overweight men [mean age 51.5 yr, body mass index (BMI) 27.6 kg·m-2] for 4 wk. In HFD-fed mice, Totum-63 reduced body weight and fat mass gain, whereas lean mass was unchanged. Moreover, fecal energy excretion was higher in Totum-63-supplemented mice, suggesting a reduction of calorie absorption in the digestive tract. In the gut, metagenomic analyses of fecal microbiota revealed a partial restoration of HFD-induced microbial imbalance, as shown by principal coordinate analysis of microbiota composition. HFD-induced increase in HOMA-IR score was delayed in supplemented mice, and insulin response to an oral glucose tolerance test was significantly reduced, suggesting that Totum-63 may prevent HFD-related impairments in glucose homeostasis. Interestingly, these improvements could be linked to restored insulin signaling in subcutaneous adipose tissue and soleus muscle. In the liver, HFD-induced steatosis was reduced by 40% (as shown by triglyceride content). In the subsequent study in men, Totum-63 (5 g·day-1) improved glucose and insulin responses to a high-carbohydrate breakfast test (84% kcal carbohydrates). It was well tolerated, with no clinically significant adverse events reported. Collectively, these data suggest that Totum-63 could improve glucose homeostasis in both HFD-fed mice and overweight individuals, presumably through a multitargeted action on different metabolic organs.NEW & NOTEWORTHY Totum-63 is a novel polyphenol-rich natural composition developed to reduce the risk of T2D. Totum-63 showed beneficial effects on glucose homeostasis in HFD-fed mice, presumably through a multitargeted action on different metabolic organs. Totum-63 was well tolerated in humans and improved postprandial glucose and insulin responses to a high-carbohydrate breakfast test.

Optimization, Validation and Application of HPLC-PDA Methods for Quantification of Triterpenoids in Vaccinium vitis-idaea L.

Triterpenoids have regained much attention as promising multi-targeting bioactive agents of natural origin in the treatment of numerous disorders. Due to the high potential for phytopharmaceutical development, accurate qualitative and quantitative analysis of triterpenoids for screening and quality control is required. Vaccinium vitis-idaea L. (lingonberry) raw materials have aroused interest as a rich source of triterpenoids. However, currently, no validated, rapid, and easy-to-perform quantification method is available for the routine control of these compounds in lingonberries. This research aimed at developing and validating HPLC-PDA methods for the determination and screening of triterpenoids in extracts of lingonberry leaves, fruits, and flowers. The developed methods were deemed satisfactory by validation, which revealed acceptable analytical specificity, linearity (r2 > 0.9999), precision (RSD < 2%), trueness (94.70-105.81%), and sensitivity (LOD: 0.08-0.65 µg/mL). The real sample analysis demonstrated established methods applicability for quantification of 13 triterpenoids in lingonberries and emphasized differences between raw materials. Lingonberry fruits were distinguished by the richness of ursolic acid; lingonberry flowers by similar profile to fruits, but low content of neutral triterpenoids; whereas lingonberry leaves by the particularly high level of α-amyrin. Thus, the proposed methods proved to be reliable and applicable for quantification and routine analysis of triterpenoids in lingonberry samples.

Vaccinium bracteatum Thunb. as a promising resource of bioactive compounds with health benefits: An updated review.

Vaccinium bracteatum Thunb.(VBT) is well-known for many physiological and bioactivities in some ancient Chinese pharmacopeias and modern researches. The health benefits are related to the presence of various nutritional and bioactive compounds. This review aims to demonstrate an updated overview of VBT in respect of botanical characters, nutritional and bioactive composition, main biological activities, and current applications. Various studies have emphasized at promising health benefits of VBT against hyperglycemia, oxidative stress, inflammation, depressive disorder, and retinal damage. However, the applications of VBT are limited to some native traditional foods and Chinese medicine. The novel beneficial efficacy and applications are still needed to be investigated. In conclusion, more research is necessary to overcome these gaps between the in-depth insights of health benefits and potential industrial applications. This review will contribute in future research for developing the functional foods derived from VBT.

Antimicrobial Photodynamic Treatment with Mother Juices and Their Single Compounds as Photosensitizers.

The potent antimicrobial effects of antimicrobial photodynamic therapy (aPDT) with visible light plus water-filtered infrared-A irradiation and natural compounds as photosensitizers (PSs) have recently been demonstrated. The aim of this study was to obtain information on the antimicrobial effects of aPDT with mother juices against typical cariogenic oral Streptococcus pathogens in their planktonic form and determine its eradication potential on total human salivary bacteria from volunteers. Mother juices of pomegranate, bilberry, and chokeberry at different concentrations were used as PSs. The unweighted (absolute) irradiance was 200 mW cm-2, applied five minutes. Planktonic cultures of Streptococcus mutans and Streptococcus sobrinus and total mixed bacteria from pooled saliva of volunteers were treated with aPDT. Up to more than 5 log10 of S. mutans and S. sobrinus were killed by aPDT with 0.4% and 0.8% pomegranate juice, 3% and 50% chokeberry juice, and 12.5% bilberry juice (both strains). Concentrations of at least 25% (pomegranate) and >50% (chokeberry and bilberry) eradicated the mixed bacteria in saliva samples. This pilot study has shown that pomegranate mother juice is superior to the berry juices as a multicomponent PS for killing pathogenic oral bacteria with aPDT.

Analysis of composition, morphology, and biosynthesis of cuticular wax in wild type bilberry (Vaccinium myrtillus L.) and its glossy mutant.

In this study, cuticular wax load, its chemical composition, and biosynthesis, was studied during development of wild type (WT) bilberry fruit and its natural glossy type (GT) mutant. GT fruit cuticular wax load was comparable with WT fruits. In both, the proportion of triterpenoids decreased during fruit development concomitant with increasing proportions of total aliphatic compounds. In GT fruit, a higher proportion of triterpenoids in cuticular wax was accompanied by a lower proportion of fatty acids and ketones compared to WT fruit as well as lower density of crystalloid structures on berry surfaces. Our results suggest that the glossy phenotype could be caused by the absence of rod-like structures in GT fruit associated with reduction in proportions of ketones and fatty acids in the cuticular wax. Especially CER26-like, FAR2, CER3-like, LTP, MIXTA, and BAS genes showed fruit skin preferential expression patterns indicating their role in cuticular wax biosynthesis and secretion.

Impact of short-term bilberry supplementation on glycemic control, cardiovascular disease risk factors, and antioxidant status in Chinese patients with type 2 diabetes.

Bilberry (Vaccinium myrtillus L.) is one of the richest natural sources of anthocyanins which are powerful antioxidants and reported to have antiinflammatory, antidyslipidemic, antihypertensive, and hypoglycemic effects. The objective of this study was to assess the effect of bilberry supplementation on biomarkers of glycemic control, lipid profile, antioxidant, and inflammatory status in patients with type 2 diabetes in a randomized, double-blind, placebo-controlled cross-over study. Twenty patients were randomized to receive either bilberry supplementation (1.4 g/day of extract) daily for 4 weeks followed by 6 weeks of washout and then an additional 4 weeks of matching placebo or vice versa. Blood pressure, metabolic parameters, antioxidant status, and oxidative stress were measured before and after each period. Results showed no effect on body weight, blood pressure, or lipid profile. HbA1c was reduced by 0.31 ± 0.58% during bilberry supplementation, but this change was not significantly different from that with placebo. Antioxidant status, oxidative stress, and inflammatory status showed no significant differences across treatments. This short-term study of bilberry supplementation did not show significant effects on cardiovascular risk factors or antioxidant status, but the tendency for improved glycemic control may suggest a longer treatment period may be effective in diabetic patients.

A simple and fast method for metabolomic analysis by gas liquid chromatography-mass spectrometry.

INTRODUCTION: The metabolomic profile is an essential tool for understanding the physiological processes of biological samples and their changes. In addition, it makes it possible to find new substances with industrial applications or use as drugs. As GC-MS is a very common tool for obtaining the metabolomic profile, a simple and fast method for sample preparation is required. OBJECTIVES: The aim of this research was to develop a direct derivatization method for GC-MS to simplify the sample preparation process and apply it to a wide range of samples for non-targeted metabolomic analysis purposes. METHODS: One pot combined esterification of carboxylic acids with methanol and silylation of the hydroxyl groups was achieved using a molar excess of chlorotrimethylsilane with respect to methanol in the presence of pyridine. RESULTS: The metabolome profile obtained from different samples, such as bilberry and cherry cuticles, olive leaves, P. aeruginosa and E. coli bacteria, A. niger fungi and human sebum from the ceruminous gland, shows that the procedure allows the identification of a wide variety of metabolites. Aliphatic fatty acids, hydroxyfatty acids, phenolic and other aromatic compounds, fatty alcohols, fatty aldehydes dimethylacetals, hydrocarbons, terpenoids, sterols and carbohydrates were identified at different MSI levels using their mass spectra. CONCLUSION: The metabolomic profile of different biological samples can be easily obtained by GC-MS using an efficient simultaneous esterification-silylation reaction. The derivatization method can be carried out in a short time in the same injection vial with a small amount of reagents.

Plant growth shapes the effects of elevation on the content and variability of flavonoids in subalpine bilberry stands.

The study of morphological and physiological responses of shrubs to climate is crucial for the understanding of future scenarios regarding climate change. In this light, studying shrub growth and physiological acclimation along an elevation gradient might be insightful. The phenolic metabolic pathway represents a powerful tool to interpret such processes. In the South-Eastern Alps, we investigated the relationships between elevation, plant traits (i.e. age, xylem ring width, annual shoot length), plant-plant interaction (i.e. shrub cover) and flavonoids in Vaccinium myrtillus L. (leaves, berries) in stands above the treeline. The relationships were parsed within causal networks using a confirmatory path analysis. Elevation was the main driver of V. myrtillus growth, having both direct and indirect effects on the leaf flavonoid content, but this was less evident for berries. In particular, the content of foliar flavonoids showed a peak at mid-elevation and where the growth of xylem rings was intermediate, while it decreased in stands with higher shoot length. Flavonoid content variability of both leaves and berries was affected by elevation and shoot length. In berries, flavonoid variability was further related to all growth traits and shrub cover. These findings evidence that flavonoid content is influenced by both elevation and growth traits of V. myrtillus, often showing non-linear relationships. These results suggest a trait-mediated response of this plant to climate conditions as a result of trade-offs between plant growth, plant defence, environmental stress and nutrient/resource availability.