Bilberry (Vaccinium myrtillus L.) fruits are an important part of local diets in many countries and are used as a medicinal herb to treat various disorders. Extracts from fruits are often a part of eye health-promoting supplements, whereas extracts from leaves are advertised for type 2 diabetes mellitus and glycemic control. This review provides an overview of the current knowledge of the phytochemical contents of bilberry fruits and leaves and their bioactivities, critically summarizes origins of the health claims and the outcome of clinical trials, with special attention towards those published in the past 10 years. Overall, the three most referenced indications, which are type 2 diabetes mellitus, vision disorders and circulatory diseases, all include contradictory results with no clear conclusion as to the benefits and recommended dosages. Moreover, the indications for vision disorders and diabetes originate from unproven or false claims that have been repeated in research since the 20^th century without consistent fact-checking. Beneficial clinical results have been attested for the treatment of dyslipidemia and chronic inflammatory disorders when applied as dietary supplementation of fresh bilberries or as anthocyanin-rich bilberry fruit extracts. However, there is a general lack of double-blinded controlled research with larger sample sizes.

The berries of Vaccinium myrtillus L. are usually collected in the wild for the purpose of being a food source. They are naturally high in phenolic compounds, which possess antioxidative properties, so the berries are therefore often labeled as “functional foods”. This study evaluated seven samples of bilberry fruits from different locations in Slovakia for the content of the main phenolic compounds (anthocyanins, flavonoids and tannins) using European Pharmacopoeia 9 spectrophotometric methods. A thorough analysis of environmental factors showed that several phenolic constituents are closely corresponding with their respective environments, as well as with each other. The environmental factors with statistically significant correlations in this study are altitude, habitat type, sunlight exposure, and soil carbon content. Our findings suggest that the berries collected at sunny sites with no topsoil damage contain more phenolic compounds. The lowest amounts of phenolic compounds were found in samples from dense forests or with visible soil erosion and windthrow damage. The negative effect of windthrow damage on the levels of secondary metabolites in bilberry fruits has been described for the first time. This study observed no relationship between the amount of phenolic compounds and soil pH, soil nitrogen levels, or slope exposition.

The aim of this study was to analyze the binding interactions between a common antihypertensive drug (amlodipine besylate—AML) and the widely distributed plant flavonoid quercetin (Q), in the presence of human serum albumin (HSA). Fluorescence analysis was implemented to investigate the effect of ligands on albumin intrinsic fluorescence and to define the binding and quenching properties. Further methods, such as circular dichroism and FT-IR, were used to obtain more details. The data show that both of these compounds bind to Sudlow’s Site 1 on HSA and that there exists a competitive interaction between them. Q is able to displace AML from its binding site and the presence of AML makes it easier for Q to bind. AML binds with the lower affinity and if the binding site is already occupied by Q, it binds to the secondary binding site inside the same hydrophobic pocket of Sudlow’s Site 1, with exactly the same affinity. Experimental data were complemented with molecular docking studies. The obtained results provide useful information about possible pharmacokinetic interactions upon simultaneous co-administration of the food/dietary supplement and the antihypertensive drug.

The aim of this study was to analyze the binding interactions between a common antihypertensive drug (ramipril, R) and the widely distributed plant flavonoid quercetin (Q), in the presence of human serum albumin (HSA). From the observed fluorescence spectra of the (HSA + R) system we can assume that ramipril is also one of the Site 3 ligands—similar to fusidic acid—the binding of which has been proven by RTG crystallography. Our claim is supported by near-UV CD spectroscopy, microscale themophoresis and molecular modeling. The presence of R slightly inhibited the subsequent binding of Q to HSA and, on the contrary, the pre-incubation of HSA with Q caused a stronger binding of R, most likely due to allosteric interactions. At high concentrations, R is also able to displace Q from its binding site. The dissociation constant K_D for the binding of R is more than hundredfold larger than for Q, which means that R is a very weak binder to HSA. The knowledge of qualitative and quantitative parameters of R, as well as the methods used in this study, are important for future research into HSA binding. This study shows the importance of implementing other methods for K_D determination. Microscale thermophoresis has proved to be a novel, practical and accurate method for K_D determination on HSA, especially in cases when fluorescence spectroscopy is unable to produce usable results.

Bog bilberry (Vaccinium uliginosum L.) is a wild-growing berry native to all circumboreal regions. There is however a significant discrepancy in the uses of bog bilberry fruits around the world. There exists a strong prejudice against the use of these berries in many European countries as well as a few incidences of poisoning reported between 1906 and 1944. In Asia and North America, this fear is completely absent from the local knowledge and the bog bilberry is valued as an excellent food and medicinal plant. There has been a lack of research on the topic in the last 50 years and thus the presumed toxicity remains unproven. This review aims to gather the conflicting information from all regions where bog bilberry grows and present them in a critical way to elucidate the possible explanations for the discrepancies. There are several possible explanations for the alleged toxicity of the bog bilberry, including a fungal infection of the fruits, individual intolerance or accidental poisoning by a different plant species; the local names meaning “drunk, inebriating, vomit-inducing berry” may be related to the alcoholic drinks made from them. This review highlights the gap in knowledge and serves as a theoretical framework for future research.

Vaccinium uliginosum (bog bilberry) is widely consumed in North America and Asia but has been historically avoided in many parts of Europe due to its alleged poisonous effects. We aimed to address this discrepancy in a systematic way with a combined phytochemical and ethnopharmacological approach, using UHPLC and UHPSFC for the chemical analysis, model organisms Caenorhabditis elegans and human liver cells GFP-Huh-7 for the bioactivity and toxicity testing, as well as fermentation experiments. Phytochemical analysis revealed minimal differences in the metabolite pattern between European and North American samples, with no evidence of toxic alkaloids or harmful secondary metabolites. Extracts exhibited no strongly toxic effects in the tested concentrations, neither in vitro (cell viability) nor in vivo (C. elegans). Berries infected by Monilinia megalospora showed altered flavonoid and anthocyanin contents but no increased toxicity. Notably, bog bilberries demonstrated a fermentation potential superior to Vaccinium myrtillus, resulting in an alcohol content of 4.8–5.8% ABV in unsweetened juices, thus potentially explaining historical accounts of inebriation. In conclusion, direct toxicity derived from these fruits is unlikely, but the alcohol content due to fruit fermentation is a plausible explanation for the folklore names (“drunk, inebriating berry”). However, additional factors such as human error, individual intolerance, or endophytic activity need to be considered.