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The aim of the study was to design microcapsules with a core of blueberry fruit extracts (Vaccinium myrtillus L.) using the ionotropic gelation method and then assess the effect of the type of extracts used and the combination of polymers on the profile of phenolic compounds, their in vitro bioavailability, stability during storage, as well as their antioxidant characteristics and cytotoxic activity against colon cancer cells while assessing biocompatibility against normal colon epithelial cells. Encapsulation efficiency (EE), ranging from 8.79 to 74.55%, significantly depended on the extract used and the type of carrier. It was shown that the addition of pectin (Pect) and whey protein isolate (WPI) to alginate (Alg) improved the efficiency of the encapsulation process. For this version of microcapsules, the highest antioxidant activity, phenolic compound content and their stability during storage were also demonstrated. The estimated content of phenolic compounds ranged from 0.48 to 40.07 mg/g, and the dominant compound was cyanidin 3-O-glucoside. In turn, the highest bioavailability of these compounds and the highest cytotoxic activity against cancer cells were characterized by microcapsules with Alg and WPI. Nevertheless, good biocompatibility with normal colon epithelial cells was demonstrated for all versions of microcapsules. The obtained data indicate that the tested variants of microcapsules protect the bioactive compounds of blueberry fruit extracts, which translates into maintaining their health-promoting properties.

Medicinal plants, through their complex chemical composition, have therapeutic properties, they can act preventively or curatively in alleviating or even treating various ailments. Both mistletoe (Viscum album L.) and blueberry (Vaccinium myrtillus L) are medicinal plants rich in polyphenols, such as flavonoids, hydroxybenzoic acid, derivatives of hydroxycinnamic acids. In the present research, the bioactive compounds were firstly extracted from mistletoe leaves (Viscum album L.) harvested from acacia (Robinia pseudoacacia L.) and blueberry fruits (Vaccinium myrtillus L.) using ethanol as solvent in two concentrations (25 and 50 %) The total phenols content, respectively the antioxidant capacity recorded by the FRAP assay, were recorded from both of the individual hydroalcoholic extracts of mistletoe and blueberry and of the combined ones, mistletoe and blueberry. Mistletoe leaves and blueberry fruits contain a considerable amount of phenolic compounds, and our results suggested that their combination can lead to obtaining a functional beverage with a high antioxidant capacity with a beneficial effect on human health.

Isolation-by-distance (IBD) pattern among bilberry (Vaccinium myrtillus L.) populations has previously been reported for this species in northern Europe. However, the number of molecular studies conducted on bilberry, using everything from isoenzymes, RAPDs to microsatellite markers, are very few and far between. Considering that Bosnia and Herzegovina (B&H) is a country rich with diverse fruit genetic resources, conducting a genetic characterization of the naturally occurring V. myrtillus populations could yield valuable data for the conservation and utilization of this resource. This study entailed genotyping samples collected from three bilberry populations located in Fojnica, Kladanj, and Srebrenica municipalities using seven polymorphic microsatellite or SSR (simple sequence repeats) markers. The obtained molecular data was used to calculate the correlation between the physical distance of the individual B&H populations and a parameter of the genetic differentiation (pairwise Fst). The results of the correlation analyses revealed an absence of a significant isolation-by-distance pattern among the three examined B&H bilberry populations. In addition, the most pronounced genetic differentiation was detected between the Srebrenica and each of the two remaining B&H populations. At the same time, the values for pFst were significant, albeit much lower, between the Fojnica and Kladanj populations. Bilberries from the sampled Srebrenica population appear to be distinct from the other B&H populations, possibly due to the different genetic origin of this population.

This study explores the bioactive potential of young shoots from blackcurrant, European blueberry, and mountain cranberry, widely employed in gemmotherapy and phytotherapy, as rich sources of antioxidants, antimicrobial agents, and anti-inflammatory components. The primary aims of this study were to enhance the extraction conditions for bioactive compounds from blackcurrant young shoots using Modde software for experimental design, to conduct a comprehensive phytochemical analysis of blackcurrant, European blueberry, and mountain cranberry young shoot extracts through LC–MS analysis, and to evaluate the in vitro biological activities of these optimized extracts. The experimental design comprised multiple variables: extraction techniques, solvent type, extraction time, apparent pH, and the solvent-to-vegetal product ratio. The responses included total phenolic content, total flavonoid content, condensed tannin content, and total antioxidant activity determined through the DPPH assay. Furthermore, the antioxidant potential of the extracts was validated through in vitro cell culture experiments, in addition to the cytotoxicity assessments conducted on both normal and cancer cell lines. Extracts obtained through Ultra-Turrax extraction using 70% acetone displayed high levels of polyphenolic compounds and enhanced antioxidant potential, regardless of young shoots origin. LC–MS analysis revealed the predominant occurrence of chlorogenic acid, hyperoside, and isoquercitrin in all examined samples. The optimized extracts also displayed significant biological potential when evaluated in vitro on cell lines. These results provide valuable insights into the potent bioactive components present in these young shoot extracts, paving the way for further exploration in therapeutic applications.

WRKY transcription factor genes compose an important family of transcriptional regulators that are present in several plant species. According to previous studies, these genes can also perform important roles in bilberry (Vaccinium myrtillus L.) metabolism, making it essential to deepen our understanding of fruit ripening regulation and anthocyanin biosynthesis. In this context, the detailed characterization of these proteins will provide a comprehensive view of the functional features of VmWRKY genes in different plant organs and in response to different intensities of light. In this study, the investigation of the complete genome of the bilberry identified 76 VmWRKY genes that were evaluated and distributed in all twelve chromosomes. The proteins encoded by these genes were classified into four groups (I, II, III, and IV) based on their conserved domains and zinc finger domain types. Fifteen pairs of VmWRKY genes in segmental duplication and four pairs in tandem duplication were detected. A cis element analysis showed that all promoters of the VmWRKY genes contain at least one potential cis stress-response element. Differential expression analysis of RNA-seq data revealed that VmWRKY genes from bilberry show preferential or specific expression in samples. These findings provide an overview of the functional characterization of these proteins in bilberry.

The aim of this study was to determine the activity of post-Chernobyl 137Cs in Slovak bilberry (Vaccinium myrtillus L.) from selected locations of Slovakia depending upon: the season, location and the soil-plant transfer factor (TF). The 137Cs activity was determined in samples of forest soil and bilberry bushes (stems, leaves and fruits) growing on the soil in three locations: Arboretum Mlyňany (1), Hodruša-Hámre (2) and Javorníky (3). Altogether we collected 35 samples; namely 15 samples from Arboretum Mlyňany, 10 samples from Hodruša-Hámre and 10 from Javorníky. The samples of soil were examined also for the activity of 40K. The samples were collected in the spring, summer and autumn in the years 2019 and 2020 and were analysed by a gamma-ray spectrometry method. The highest levels of 137Cs activity concentrations were determined in Hodruša-Hámre in September where they reached 161 ± 13 Bq.kg−1 in the soil, 3.95 ± 1.07 Bq.kg−1 in the stems and 14.7 ± 4.26 Bq.kg−1 in the leaves. The lowest levels were determined in the Arboretum Mlyňany in October where the 137Cs activity in the soil reached 13.1 ± 1 15 Bq.kg−1, while in the stems, leaves and fruits it was lower than the minimum detectable level. In the latter location, we also determined the highest activity of 40K, the radioactive isotope physiologically and metabolically similar to 137Cs. The mass activity of 137Cs in all samples of bilberries were lower than the minimum detectable activity (MDA). The comparison of the results obtained in this study with the limit for total mass activity of radiocaesium (134Cs + 137Cs) in the food (600 Bq.kg−1) allowed us to conclude that the fruits, stems and leaves of Slovak bil-berries present neither health nor environmental risk.

Background:Vaccinium myrtillus L. is a species belonging to the genus Vaccinium of the family Ericaceae. Bilberries have drawn attention due to the multiple benefits for the human health, including antioxidant, anti-inflammatory, anticancer, anti-neurodegenerative, and cardioprotective effects. Recently, bilberries were shown to inhibit the activity of carbohydrate-hydrolysing enzymes that can help reduce the intensity of the metabolic syndrome and prevent type 2 diabetes.

The purpose of the current study was to identify and quantify triterpenoids and other non-polar compounds in the leaves of three high bush blueberry cultivars (Vaccinium corymbosum L. var. Bluegold, var. Bluecrop and var. Elliott) and three natural populations of Vaccinium species (Vaccinium uliginosum L., Vaccinium myrtillus L. and Vaccinium vitis-idaea L.) by means of gas chromatography mass spectrometry (GC-MS) and high-performance liquid chromatography with diode array detector (HPLC-DAD). Metabolite profiles differed significantly among the Vaccinium species analyzed, as well as among the populations of the same species. The populations of V. vitis-idaea predominantly contained relative concentrations of phytosterols (varying between 10.48% of total ion current (TIC) and 22.29% of TIC) and almost twice the content of triterpenes (from 29.84% of TIC to 49.62% of TIC) of the other berry species investigated. The leaves of V. corymbosum varieties biosynthesized the highest relative amount of fatty acids, while the leaves of the populations of V. uliginosum had the highest relative concentrations of fatty alcohols. The results of principal component analysis (PCA) and hierarchical cluster analysis (HCA) showed that the diverse populations of each berry species analyzed differed from each other, most likely due to variations in the climatic and geographical conditions of their localities.

Bilberry is a valuable wild forest fruit harvested in many countries in Europe. The biological activities of bilberry include antioxidant, anticancer, antiviral, antibacterial, and anticholinesterase activities. This study examines the protective effects of a bilberry (BB) preparation on IEC-6, Caco-2, and HepG2 cell lines. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to study the cytotoxicity of BB. The genotoxicity was determined using single-cell microgel electrophoresis. The Ames test was employed to assay bilberry mutagenicity. No significant effects of BB (12.5-100 µg dry mass/ml) were observed on the viability of IEC-6, Caco-2, and HepG2 cells. There were no differences in the percentage of DNA in the comet tail between the cells treated with BB (100 µg dry mass/ml) and the control cells. However, a significant reduction of oxidative DNA damage in the HepG2 cells was found. BB exhibited neither mutagenic nor promutagenic effects. Our results suggest that bilberry can be a potential tool in the prevention of chronic diseases, without any undesired effects on the cells of the gastrointestinal tract.

Anthocyanins have been associated with several health benefits, although the responsible mechanisms are not well established yet. In the present study, an anthocyanin-rich extract from bilberry (Vaccinium myrtillus L.) was tested in order to evaluate its capacity to modulate reactive oxygen species (ROS) production and resistance to thermally induced oxidative stress, using the nematode Caenorhabditis elegans as an in vivo model. The assays were carried out with the wild-type N2 strain and the mutant strains daf-16(mu86) I and hsf-1(sy441), which were grown in the presence of two anthocyanin extract concentrations (5 and 10 μg/mL in the culture medium) and further subjected to thermal stress. The treatment with the anthocyanin extract at 5 μg/mL showed protective effects on the accumulation of ROS and increased thermal resistance in C. elegans, both in stressed and non-stressed young and aged worms. However, detrimental effects were observed in nematodes treated with 10 μg/mL, leading to a higher worm mortality rate compared to controls, which was interpreted as a hormetic response. These findings suggested that the effects of the bilberry extract on C. elegans might not rely on its direct antioxidant capacity, but other mechanisms could also be involved. Additional assays were performed in two mutant strains with loss-of-function for DAF-16 (abnormal DAuer Formation factor 16) and HSF-1 (Heat Shock Factor 1) transcription factors, which act downstream of the insulin/insulin like growth factor-1 (IGF-1) signaling pathway. The results indicated that the modulation of these factors could be behind the improvement in the resistance against thermal stress produced by bilberry anthocyanins in young individuals, whereas they do not totally explain the effects produced in worms in the post-reproductive development stage. Further experiments are needed to continue uncovering the mechanisms behind the biological effects of anthocyanins in living organisms, as well as to establish whether they fall within the hormesis concept.

Kuklova M., Kukla J.: Transfer of risk elements in soil-bilberry system. Ekologia (Bratislava), Vol. 32, No. 2, p. 211-219, 2013.