On the effect of flavonoids and dietary fibre in lingonberries on atherosclerotic plaques, lipid profiles and gut microbiota composition in Apoe-/- mice.

It has not been clarified whether the anti-atherosclerotic effect of lingonberry can be ascribed to its content of flavonoids or dietary fibre or both. The aim of this study was to evaluate the metabolic effects of whole lingonberries compared with isolated flavonoid and fibre fractions on atherosclerotic plaques, plasma lipid profiles, gut microbiota and microbiota-dependent metabolites in an Apoe-/- mouse model. Mice fed whole lingonberries showed the lowest amount of atherosclerotic plaques, while mice fed the fibre fraction had the highest formation of caecal butyric acid. Flavonoids, rather than dietary fibre, were suggested to be the components that favour proliferation of Akkermansia, as judged by the lowest abundance of this bacterium in mice fed the fibre fraction. All groups fed lingonberry diets had both, lower Firmicutes/Bacteroidetes ratios and creatinine concentrations, compared with the control. To conclude, different components in lingonberries are associated with different physiological effects in Apoe-/- mice.

Aqueous extracts of lingonberry and blackberry leaves identified by high-content screening beneficially act on cholesterol metabolism.

Decreasing circulating low-density lipoprotein (LDL) cholesterol levels leads to decreased risk of cardiovascular diseases. Natural compounds are capable of lowering LDL-cholesterol even on top of lifestyle modification or medication. To identify novel plant-derived compounds to lower plasma LDL cholesterol levels, we performed high-content screening based on the transcriptional activation of the promoter of the LDL receptor (LDLR). The identified hits were thoroughly validated in human hepatic cell lines in terms of increasing LDLR mRNA and protein levels, lowering cellular cholesterol levels and increasing cellular LDL uptake. By means of this incremental validation process in vitro, aqueous extracts prepared from leaves of lingonberries (Vaccinium vitis-idaea) as well as blackberries (Rubus fruticosus) were found to have effects comparable to lovastatin, a prototypic cholesterol-lowering drug. When applied in vivo in mice, both extracts induced subtle increases in hepatic LDLR expression. In addition, a significant increase in high-density lipoprotein (HDL) cholesterol was observed. Taken together, aqueous extracts from lingonberry or blackberry leaves were identified and characterized as strong candidates to provide cardiovascular protection.

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.

Beneficial effects of lingonberry (Vaccinium vitis-idaea L.) supplementation on metabolic and inflammatory adverse effects induced by high-fat diet in a mouse model of obesity.

Obesity is a constantly increasing health problem worldwide. It is associated with a systemic low-grade inflammation, which contributes to the development of metabolic disorders and comorbidities such as type 2 diabetes. Diet has an important role in the prevention of obesity and its adverse health effects; as a part of healthy diet, polyphenol-rich berries, such as lingonberry (Vaccinium vitis-idaea L.) have been proposed to have health-promoting effects. In the present study, we investigated the effects of lingonberry supplementation on high-fat diet induced metabolic and inflammatory changes in a mouse model of obesity. Thirty male C57BL/6N mice were divided into three groups (n = 10/group) to receive low-fat (LF), high-fat (HF) and lingonberry-supplemented high-fat (HF+LGB) diet for six weeks. Low-fat and high-fat diet contained 10% and 46% of energy from fat, respectively. Lingonberry supplementation prevented the high-fat diet induced adverse changes in blood cholesterol and glucose levels and had a moderate effect on the weight and visceral fat gain, which were 26% and 25% lower, respectively, in the lingonberry group than in the high-fat diet control group. Interestingly, lingonberry supplementation also restrained the high-fat diet induced increases in the circulating levels of the proinflammatory adipocytokine leptin (by 36%) and the inflammatory acute phase reactant serum amyloid A (SAA; by 85%). Similar beneficial effects were discovered in the hepatic expression of the inflammatory factors CXCL-14, S100A10 and SAA by lingonberry supplementation. In conclusion, the present results indicate that lingonberry supplementation significantly prevents high-fat diet induced metabolic and inflammatory changes in a murine model of obesity. The results encourage evaluation of lingonberries as a part of healthy diet against obesity and its comorbidities.

Monitoring Winter Stress Vulnerability of High-Latitude Understory Vegetation Using Intraspecific Trait Variability and Remote Sensing Approaches.

In this study, we focused on three species that have proven to be vulnerable to winter stress: Empetrum nigrum, Vaccinium vitis-idaea and Hylocomium splendens. Our objective was to determine plant traits suitable for monitoring plant stress as well as trait shifts during spring. To this end, we used a combination of active and passive handheld normalized difference vegetation index (NDVI) sensors, RGB indices derived from ordinary cameras, an optical chlorophyll and flavonol sensor (Dualex), and common plant traits that are sensitive to winter stress, i.e. height, specific leaf area (SLA). Our results indicate that NDVI is a good predictor for plant stress, as it correlates well with height (r = 0.70, p < 0.001) and chlorophyll content (r = 0.63, p < 0.001). NDVI is also related to soil depth (r = 0.45, p < 0.001) as well as to plant stress levels based on observations in the field (r = -0.60, p < 0.001). Flavonol content and SLA remained relatively stable during spring. Our results confirm a multi-method approach using NDVI data from the Sentinel-2 satellite and active near-remote sensing devices to determine the contribution of understory vegetation to the total ecosystem greenness. We identified low soil depth to be the major stressor for understory vegetation in the studied plots. The RGB indices were good proxies to detect plant stress (e.g. Channel G%: r = -0.77, p < 0.001) and showed high correlation with NDVI (r = 0.75, p < 0.001). Ordinary cameras and modified cameras with the infrared filter removed were found to perform equally well.

Cranberries versus lingonberries: A challenging authentication of similar Vaccinium fruit.

Due to unique phytochemicals contained, Vaccinum berries are known to have a number of positive health effects. In this context, lingonberries (Vaccinium vitis-idaea) are considered to be the most effective, thus finding many uses. Recently, fraud suspicion on lingonberries-based products has been reported, partial or even total replacement by less valued cranberries (Vaccinium macrocarpon) was found. In this study, metabolomic fingerprinting employing instrumental platform consisting of U-HPLC-HRMS/MS was investigated for discrimination between the two Vaccinum berries species. Methanolic extracts of 33 authentic samples from two harvest years were analyzed and chemometric evaluation was performed to identify significant marker compounds, their stability during drying process was assessed, too. The characteristic markers most contributing to berries classification were representatives of polyphenols and phospholipids. Peonidin 3-O-arabinoside and myricetin 3-O-glucoside, not occurring in lingonberries, enabled to discover the presence of cranberries in prepared admixtures down to 1% (w/w).

Biochemical Properties and Neuroprotective Effects of Compounds in Various Species of Berries.

Several species of berries, such as blueberries (Vaccinium angustifolium) and lingonberries (Vaccinium vitis-idaea L.), have attracted much scientific attention in recent years, especially due to their reported antioxidant and anti-inflammatory properties. Berries, as with other types of plants, have developed metabolic mechanisms to survive various environmental stresses, some of which involve reactive oxygen species. In addition, the fruits and leaves of berries have high amounts of polyphenols, such as flavonoids, which act as potent antioxidants. These compounds could potentially be beneficial for brain aging and neurodegenerative disorders. There are now several studies documenting the beneficial effects of various berries in cell models of neurotoxicity as well as in vivo models of neurodegenerative disease. In the current review, we discuss the metabolic strategies that plants and animals have developed in order to combat reactive oxygen species. We then discuss issues of bioavailability of various compounds in mammals and provide a synopsis of studies demonstrating the neuroprotective ability of berries and polyphenols. We also summarize findings from our own research group. For example, we have detected various polyphenols in samples of blueberries and lingonberries and have found that the leaves have a much higher antioxidant capacity than the fruits. Extracts from these species have also demonstrated neuroprotective effects in cellular models of toxicity and inflammation, which are being further pursued in animal models.

Supplementing diet with Manitoba lingonberry juice reduces kidney ischemia-reperfusion injury.

BACKGROUND: Lingonberry (Vaccinium vitis-idaea L.) contains high levels of anthocyanins which are bioavailable in the kidney and may be protective against ischemia-reperfusion (IR)-induced acute kidney injury. This study investigated the effect of lingonberry juice on the IR-induced stress-activated signalling pathway and inflammatory response in the kidney. RESULTS: Sprague-Dawley rats subjected to kidney IR had significantly impaired kidney function, with increased activation of the JNK signalling pathway and increased inflammatory response, measured using a multiplex panel containing an extensive array of inflammatory biomarkers. In rats fed 1 mL lingonberry juice daily for 3 weeks prior to IR, kidney function was protected and attenuation of inflammatory response and JNK signalling was reflected in the reduction of the measured biomarkers. In vitro results in cultured HK-2 cells confirmed that lingonberry anthocyanins reduced JNK signalling and inflammatory gene expression after IR. CONCLUSION: This study shows, for the first time, that daily supplementation with lingonberry juice may protect against loss of kidney function induced by IR injury by modulating JNK signalling and inhibiting the subsequent inflammatory response. © 2017 Her Majesty the Queen in Right of Canada. Journal of the Science of Food and Agriculture © 2017 Society of Chemical Industry.

Impact of Procyanidins from Different Berries on Caspase 8 Activation in Colon Cancer.

SCOPE: The aim of this work is to identify which proapoptotic pathway is induced in human colon cancer cell lines, in contact with proanthocyanidins extracted from various berries. METHODS AND RESULTS: Proanthocyanidins (Pcys) extracted from 11 berry species are monitored for proapoptotic activities on two related human colon cancer cell lines: SW480-TRAIL-sensitive and SW620-TRAIL-resistant. Apoptosis induction is monitored by cell surface phosphatidylserine (PS) detection. Lowbush blueberry extract triggers the strongest activity. When tested on the human monocytic cell line THP-1, blueberry Pcys are less effective for PS externalisation and DNA fragmentation is absent, highlighting a specificity of apoptosis induction in gut cells. In Pcys-treated gut cell lines, caspase 8 (apoptosis extrinsic pathway) but not caspase 9 (apoptosis intrinsic pathway) is activated after 3 hours through P38 phosphorylation (90 min), emphasizing the potency of lowbush blueberry Pcys to eradicate gut TRAIL-resistant cancer cells. CONCLUSION: We highlight here that berries Pcys, especially lowbush blueberry Pcys, are of putative interest for nutritional chemoprevention of colorectal cancer in view of their apoptosis induction in a human colorectal cancer cell lines.

Influence of polyphenols from lingonberry, cranberry, and red grape on in vitro digestibility of rice.

Dietary polyphenols are abundant antioxidants in the human diet and are associated with lower rates of diabetes and cardiovascular disease. This study aims to determine the effects of cooking white rice (WR) added with lingonberry (WRLB), cranberry (WRCB), and red grape (WRRG) on in vitro digestibility. There was significantly lower level of glucose release for WRRG compared with WR (p < 0.05). WRLB and WRCB showed no effect on glucose release compared with WR (p > 0.05). Increasing concentrations of red grape polyphenol decreased digestibility of white rice (p < 0.05). A positive correlation between the red grape phenolic content and the resistant starch was observed (R = 0.9854). Red grape polyphenol had the greatest impact on reducing in vitro digestibility of white rice. The addition of polyphenols in carbohydrate-rich foods may be a practical means to reduce the high glycemic response of rice eaten around the world.

Characterization of in vitro antifungal activities of small and American cranberry (Vaccinium oxycoccos L. and V. macrocarpon Aiton) and lingonberry (Vaccinium vitis-idaea L.) concentrates in sugar reduced fruit spreads.

In this study, cranberry and lingonberry concentrates were added to commercial sugar-reduced fruit spreads (raspberry-Aloe vera, strawberry-guava, and strawberry-lime), and tested for their antifungal activities. Selected strains of the species Absidia glauca, Penicillium brevicompactum, Saccharomyces cerevisiae and Zygosaccharomyces bailii, as well as xerophilic environmental isolates of the genera Penicillium and Eurotium were used for challenge testing. Initially, varying concentrations of synthetic antifungal agents, such as sodium benzoate, potassium sorbate and butyl 4-hydroxybenzoate were tested against these fungi on wort agar containing 31% fructose at different pH values. Subsequently, the experiments were conducted in fruit spreads containing different concentrations of cranberry and lingonberry concentrates. The results of this study demonstrate that these concentrates were able to inhibit growth of visible colonies of xerophilic and non-xerophilic fungi. Cranberry and lingonberry concentrates are interesting candidates for natural preservation against fungal growth in sugar reduced fruit spreads.

Dwarf shrub hydraulics: two Vaccinium species (Vaccinium myrtillus, Vaccinium vitis-idaea) of the European Alps compared.

Vaccinium myrtillus and Vaccinium vitis-idaea are two dwarf shrubs widespread in the European Alps. We studied the hydraulics of these species hypothesizing that (1) the hydraulic architecture of dwarf shrubs differs from trees, (2) hydraulic properties reflect the species' ecological amplitude and (3) hydraulic properties vary spatially and seasonally. Key hydraulic parameters (osmotic potential, turgor loss point, xylem hydraulic conductivity, vulnerability to drought-induced embolism, stomata closure, drought-induced cell damage and embolism repair) and related wood anatomical traits (conduit diameter and conduit wall reinforcement) were analyzed at four sites in Tyrol, Austria. Both species exhibited low hydraulic safety as well as low hydraulic efficiency. Fifty percentage embolism accumulated at -2.08 (V. myrtillus) and -1.97 MPa (V. vitis-idaea), 88% stomata closure was at -2.19 and -2.35 MPa, respectively. After drought, both species showed embolism repair on re-watering. Site-specific variation within species was low, while seasonal changes in embolism resistance and turgor loss point were observed. Results indicate that studied Vaccinium species have a high risk for embolism formation. This is balanced by refilling capacities, which are probably based on the small growth height of dwarf shrubs. V. vitis-idaea, which occurs on drier sites, showed more efficient repair and a lower turgor loss point than V. myrtillus.

Characterization of metabolite profiles of leaves of bilberry (Vaccinium myrtillus L.) and lingonberry (Vaccinium vitis-idaea L.).

Leaves of bilberry (Vaccinium myrtillus L.) and lingonberry (Vaccinium vitis-idaea L.) are potential raw materials for food and health care products. Targeted (HPLC-DAD, HPLC-MS, and GC-FID) and nontargeted ((1)H NMR) approaches were applied to study the metabolomic profiles of these leaves. Chlorogenic acid was the major phenolic compound in bilberry leaves and arbutin in lingonberry leaves. Flavonol glycosides were another major group of phenolics in bilberry [5-28 mg/g DM (dry mass)] and lingonberry (15-20 mg/g DM) leaves. Contents of fatty acids were analyzed using GC-FID. The changes in the metabolomics profile during the season were apparent in bilberry but not lingonberry leaves. Negative correlation was found between the contents of lipids and phenolics. The consistency between the key results obtained by targeted and nontargeted analyses suggests nontargeted metabolomic analysis is an efficient tool for fast screening of various leaf materials.

1H NMR-based metabolic fingerprinting of urine metabolites after consumption of lingonberries (Vaccinium vitis-idaea) with a high-fat meal.

The use of NMR metabolomics in clinical trials is growing; however, reports of postprandial experiments in humans are scarce. The present study investigated whether consumption of lingonberries as a supplement to an oil-rich meal modifies the postprandial fingerprints of human urine. Urine samples were analysed by (1)H NMR, and untargeted multivariate analysis was applied to the data for comprehensive fingerprinting. A clear separation of postprandial lingonberry meal samples was revealed. To evaluate statistical differences, a targeted approach was applied for the informative spectral areas. Significantly (p<0.05) increased levels of polyphenol metabolites, hippuric acid and 4-hydroxyhippuric acid, and decreased creatinine and dimethylamine levels were the major explanations for the grouping of the postprandial samples after the different meals. Thus, inclusion of polyphenol-rich lingonberry powder in a rapeseed oil-rich meal modifies the metabolic profile of urine which may be used to reveal both consumption of berries and health-promoting changes in the common metabolism.

A C-repeat binding factor transcriptional activator (CBF/DREB1) from European bilberry (Vaccinium myrtillus) induces freezing tolerance when expressed in Arabidopsis thaliana.

Freezing stress affects all plants from temperate zones to the poles. Global climate change means such freezing events are becoming less predictable. This in turn reduces the ability of plants to predict the approaching low temperatures and cold acclimate. This has consequences for crop yields and distribution of wild plant species. C-repeat binding factors (CBFs) are transcription factors previously shown to play a vital role in the acclimation process of Arabidopsis thaliana, controlling the expression of hundreds of genes whose products are necessary for freezing tolerance. Work in other plant species cements CBFs as key determinants in the trait of freezing tolerance in higher plants. To test the function of CBFs from highly freezing tolerant plants species we cloned and sequenced CBF transcription factors from three Vaccinium species (Vaccinium myrtillus, Vaccinium uliginosum and Vaccinium vitis-idaea) which we collected in the Arctic. We tested the activity of CBF transcription factors from the three Vaccinium species by producing transgenic Arabidopsis lines overexpressing them. Only the Vaccinium myrtillus CBF was able to substantially activate COR (CBF-target) gene expression in the absence of cold. Correspondingly, only the lines expressing the Vaccinium myrtillus CBF were constitutively freezing tolerant. The basis for the differences in potency of the three Vaccinium CBFs was tested by observing cellular localisation and protein levels. All three CBFs were correctly targeted to the nucleus, but Vaccinium uliginosum CBF appeared to be relatively unstable. The reasons for lack of potency for Vaccinium vitis-idaea CBF were not due to stability or targeting, and we speculate that this was due to altered transcription factor function.

Application of fungistatics in soil reduces N uptake by an arctic ericoid shrub (Vaccinium vitis-idaea).

In arctic tundra soil N is highly limiting, N mineralization is slow and organic N greatly exceeds inorganic N. We studied the effects of fungistatics (azoxystrobin [Quadris] or propiconazole [Tilt]) on the fungi isolated from ericaceous plant roots in vitro. In addition to testing the phytotoxicity of the two fungistatics we also tested their effects on growth and nitrogen uptake of an ericaceous plant (Vaccinium uliginosum) in a closed Petri plate system without root-associated fungi. Finally, to evaluate the fungistatic effects in an in vivo experiment we applied fungistatics and nitrogen isotopes to intact tundra soil cores from Toolik Lake, Alaska, and examined the ammonium-N and glycine-N use by Vaccinium vitis-idaea with and without fungistatics. The experiments on fungal pure cultures showed that Tilt was more effective in reducing fungal colony growth in vitro than Quadris, which was highly variable among the fungal strains. Laboratory experiments aiming to test the fungistatic effects on plant performance in vitro showed that neither Quadris nor Tilt affected V. uliginosum growth or N uptake. In this experiment V. uliginosum assimilated more than an order of magnitude more ammonium-N than glycine-N. The intact tundra core experiment provided contrasting results. After 10 wk of fungistatic application in the growth chamber V. vitis-idaea leaf %N was 10% lower and the amount of leaf 15N acquired was reduced from labeled ammonium (33%) and glycine (40%) during the 4 d isotope treatment. In contrast to the in vitro experiment leaf 15N assimilation from glycine was three times higher than from 15NH4 in the treatments that received no-fungistatics. We conclude that the function of the fungal communities is essential to the acquisition of N from organic sources and speculate that N acquisition from inorganic sources is mainly inhibited by competition with complex soil microbial communities.

Impacts of extreme winter warming events on plant physiology in a sub-Arctic heath community.

Insulation provided by snow cover and tolerance of freezing by physiological acclimation allows Arctic plants to survive cold winter temperatures. However, both the protection mechanisms may be lost with winter climate change, especially during extreme winter warming events where loss of snow cover from snow melt results in exposure of plants to warm temperatures and then returning extreme cold in the absence of insulating snow. These events cause considerable damage to Arctic plants, but physiological responses behind such damage remain unknown. Here, we report simulations of extreme winter warming events using infrared heating lamps and soil warming cables in a sub-Arctic heathland. During these events, we measured maximum quantum yield of photosystem II (PSII), photosynthesis, respiration, bud swelling and associated bud carbohydrate changes and lipid peroxidation to identify physiological responses during and after the winter warming events in three dwarf shrub species: Empetrum hermaphroditum, Vaccinium vitis-idaea and Vaccinium myrtillus. Winter warming increased maximum quantum yield of PSII, and photosynthesis was initiated for E. hermaphroditum and V. vitis-idaea. Bud swelling, bud carbohydrate decreases and lipid peroxidation were largest for E. hermaphroditum, whereas V. myrtillus and V. vitis-idaea showed no or less strong responses. Increased physiological activity and bud swelling suggest that sub-Arctic plants can initiate spring-like development in response to a short winter warming event. Lipid peroxidation suggests that plants experience increased winter stress. The observed differences between species in physiological responses are broadly consistent with interspecific differences in damage seen in previous studies, with E. hermaphroditum and V. myrtillus tending to be most sensitive. This suggests that initiation of spring-like development may be a major driver in the damage caused by winter warming events that are predicted to become more frequent in some regions of the Arctic and that may ultimately drive plant community shifts.

Reciprocal carbon and nitrogen transfer between an ericaceous dwarf shrub and fungi isolated from Piceirhiza bicolorata ectomycorrhizas.

The overstorey coniferous trees and understorey ericaceous dwarf shrubs of northern temperate and boreal forests have previously been considered to form mycorrhizas with taxonomically and functionally distinct groups of fungi. Here, we tested the hypothesis that Meliniomyces variabilis and Meliniomyces bicolor, isolated from Piceirhiza bicolorata ectomycorrhizas of pine, can function as ericoid mycorrhizal symbionts with Vaccinium vitis-idaea. We used split-compartment microcosms to measure the reciprocal exchange of (13)C and (15)N between V. vitis-idaea and three fungal isolates in the Hymenoscyphus ericae aggregate isolated from Scots pine ectomycorrhizas (M. variabilis and M. bicolor) or Vaccinium roots (M. variabilis). The extramatrical fungal mycelium of labelled mycorrhizal plants was significantly enriched in (13)C, and the leaves were significantly enriched in (15)N, compared with nonmycorrhizal and nonlabelled controls. * These findings show for the first time that fungi in the H. ericae aggregate, isolated from pine ectomycorrhizas, can transfer C and N and can thus form functional ericoid mycorrhizas in an understorey ericaceous shrub.