Bilberry Extract Research Articles

European bilberry extract reduces high-temperature baked food-induced accumulation of Nε-carboxymethyllysine and Nε-carboxyethyllysine in vivo

This study aimed to investigate the effect of European bilberry extract (EBE) on the accumulation of Nε-carboxymethyllysine (CML) and Nε-carboxyethyllysine (CEL) in rats exposed to a high advanced glycation end products (AGEs) diet. We found that EBE reduced high AGEs diet-induced accumulation of free-CML, bound-CML, free-CEL, and bound-CEL in the serum, kidney, skin, and brain. EBE also inhibited high AGEs diet-induced accumulation of bound-CML and bound-CEL in the uterus, ovary, stomach, duodenum, and colon. Meanwhile, EBE attenuated high AGEs diet-induced accumulation of free-CML and free-CEL in the muscle, bone, joint, and eyes. In addition, EBE ameliorated high AGEs diet-induced accumulation of free-CML and bound-CML in the liver, free-CML in the ovary, and bound-CML in the thyroid gland. EBE had no effect on the accumulation of free-CML, bound-CML, free-CEL, and bound-CEL in the adrenal gland and free-CML and free-CEL in the heart caused by a high AGEs diet. We did not observe AGEs accumulation in the pancreas, aorta, lung, spleen, and adipose tissues. This study revealed the in vivo distribution of CML and CEL exposed to a high AGEs diet and the effect of EBE on reducing the accumulation of CML and CEL in the specific target tissues.

Anthocyanin-Rich Berry Extracts Affect SN-38-Induced Response: A Comparison of Non-Tumorigenic HCEC-1CT and HCT116 Colon Carcinoma Cells.

Chemotherapy with irinotecan (CPT-11), the pro-drug of the highly cytotoxic SN-38, is among the standard-of-care treatments for colorectal cancer. To counteract undesired toxic side effects on healthy tissue such as the intestinal epithelium, the use of preparations rich in polyphenols with anti-oxidative and anti-inflammatory properties such as anthocyanins has been proposed. In the present study, the question of whether non-tumorigenic human epithelium cells (HCEC-1CT) can be protected against the cytotoxic impact of SN-38 by anthocyanin-rich polyphenol extracts without compromising the desired therapeutic effect against tumor cells (HCT-116) was addressed. Hence, single and combinatory effects of anthocyanin-rich polyphenol extracts of elderberry (EB), bilberry (Bil), blackberry (BB) and black currant (BC) with the chemotherapeutic drug SN-38 were investigated. Out of the extracts, BB showed the most potent concentration-dependent cytotoxicity alone and in combination with SN-38, with even stronger effects in non-tumorigenic HCEC-1CT cells. In cytotoxic concentrations, BB decreased the level of DNA/topoisomerase I covalent complexes in HCEC-1CT cells below base level but without concomitant reduction in SN-38-induced DNA strand breaks. The herein reported data argue towards an interference of anthocyanins with successful treatment of cancer cells and a lack of protective properties in healthy cells.

Natural product-based treatment potential for type 2 diabetes mellitus and cardiovascular disease.

Type 2 diabetes (T2D) is a metabolic disease of impaired glucose utilization and a major cause of cardiovascular disease (CVD). The pathogenesis of both diseases shares common risk factors and mechanisms, and both are significant contributors to global morbidity and mortality. Supplements of natural products for T2D mellitus (T2DM) and CVD can be seen as a potential preventive and effective therapeutic strategy. To critically evaluate the therapeutic potential of natural products in T2D and coronary artery disease (CAD). By using specific keywords, we strategically searched the PubMed database. Randomized controlled trials (RCTs) were searched as the primary focus that examined the effect of natural products on glycemic control, oxidative stress, and antioxidant levels. We focused on outcomes such as low blood glucose levels, adjustment on markers of oxidative stress and antioxidants. After screening full-length papers, we included 9 RCTs in our review that met our inclusion criteria. In the literature search on the database, we found that various natural products like plant secondary metabolites play a diverse role in the management of CAD. American ginseng, sesame oil and cocoa flavanols proved effective in lowering blood glucose levels and controlling blood pressure, which are key factors in managing T2DM and CVD. In diabetic patients Melissa officinalis effectively reduce inflammation and shows diabetes prevention. Both fish oil and flaxseed oil reduced insulin levels and inflammatory markers, suggesting benefits for both conditions. The lipid profile and endothelial function were enhanced by Nigella sativa oil and Terminalia chebula, which is significant for the management of cardiovascular risk factors in T2DM. Additionally Bilberry extract also showed promise for improving glycemic control in patients with T2DM. The high level of antioxidant, anti-inflammatory, and anti-angiogenic properties found in natural products makes them promising therapeutic options for the management of CAD, with the potential benefit of lowering the risk of CAD.

Antiaging, Brightening, and Antioxidant Efficacy of Fermented Bilberry Extract (Vaccinium myrtillus): A Randomized, Double-Blind, Placebo-Controlled Trial.

Strategies for successful aging, including the use of food supplements, are part of the approach to support skin youthfulness. To demonstrate the efficacy of fermented bilberry extract (FBE) against skin aging and uneven complexion, a clinical trial was carried out on 66 subjects with visible "crow's feet" wrinkles, mild-to-moderate skin slackness, and uneven skin tone. The wrinkle depth, skin smoothness (Ra) and roughness (Rz), skin firmness (R0) and elasticity (R2), skin coloration (ITA°), and skin antioxidant capacity were measured before and after 28 (D28), 56 (D56), and 84 (D84) days of product use (either FBE or a placebo). These parameters were also integrated with a clinical evaluation, carried out by a dermatologist, and a self-assessment questionnaire to align the measured efficacy with the visual or perceived efficacy. At D84, the wrinkle depth had decreased by 10.6%, Ra had improved by 7.9%, Rz had decreased by 7.3%, R0 had improved by 13.3%, R2 had improved by 12.4%, and skin antioxidant capacity had increased by 20.8%. ITA° increased by 20.8% and was accompanied by a decrease in the skin's redness component by 16.8% and an increase in the lightness component by 2.2%. The variation of all the above-mentioned parameters was statistically significant between the FBE and PL groups. Our findings demonstrate the efficacy of FBE in improving skin aging and complexion evenness.

Vaccinium myrtillus L. Leaf Waste as a Source of Biologically Potent Compounds: Optimization of Polyphenol Extractions, Chemical Profile, and Biological Properties of the Extracts.

The aims of the present research include (1) optimization of extraction from Vaccinium myrtillus leaf waste via investigation of plant material:medium ratio, extraction medium, and extraction period, employing extractions at room and high temperatures, or using ultrasound and microwaves (M, HAE, UAE, and MAE, respectively), (2) physicochemical characterization, and (3) investigation of extract biological potential. The statistical analysis revealed that optimal levels of parameters for the greatest polyphenolic yield were a proportion of 1:30 g/mL, ethyl alcohol 50% (v/v) during 2 min of microwave irradiation. By LC-MS analysis, 29 phenolic components were detected; HAE showed the highest richness of almost all determined polyphenols, while chlorogenic acid and quercetin 3-O-glucuronide were dominant. All extracts showed a high inhibition of Staphylococcus aureus growth. The effect of different parameters on extracts' antioxidant capacity depended on the used tests. The extracts also showed a stimulative influence on keratinocyte viability and anti-inflammatory activity (proven in cell-based ELISA and erythrocyte stabilization assays). The extraction procedure significantly affected the extraction yield (MAE ≥ maceration ≥ UAE ≥ HAE), whereas conductivity, density, surface tension, and viscosity varied in a narrow range. The presented research provides evidence on the optimal extraction conditions and technique, chemical composition, and antioxidant, antimicrobial, anti-inflammatory, and keratinocyte viability properties of bilberry extracts for potential applications in pharmacy and cosmetics.

Influence of Bilberry Extract on Neuronal Cell Toxicity.

Increased intake of dietary antioxidants such as anthocyanins, which are enriched in colourful fruits, is a promising alternative to reduce the risk of degenerative diseases such as Alzheimer's Disease (AD). Since Amyloid β (Aβ) is one of the key components contributing to AD pathology, probably by reactive oxygen species (ROS) induction, this study investigated the preventive effect of anthocyanin-rich bilberry extract (BE) and its anthocyanin fraction (ACN) on ROS generation and cell toxicity. The results showed a significant and concentration-dependent decrease in neuroblastoma cell (SH-SY5Y) viability by BE or ACN, whereas no cell toxicity was observed in HeLa cells. Incubation with BE and ACN for 24 h diminished the generation of induced ROS levels in SH-SY5Y and HeLa cells. In addition, low concentrations of BE (1-5 µg/mL) showed protective effects against Aβ-induced cytotoxicity in SH-SY5Y cells. In conclusion, our results suggest antioxidant and protective effects of BE and ACN, which could potentially be used to delay the course of neurodegenerative diseases such as AD. Further studies are needed to clarify the high potential of anthocyanins and their in vivo metabolites on neuronal function.

Potential ameliorative effects of bilberry (Vaccinium myrtillus L.) fruit extract on cisplatin-induced reproductive damage in adult male albino rats.

Cisplatin (CP) is a widely used chemotherapeutic agent, but its severe side effects impact testicular function. We investigated the potential protective effects of bilberry extract against CP-induced testicular toxicity. Forty adult male albino rats were divided into four groups. Control animals received a single oral dose of 0.9% saline. Bilberry-treated rats received oral bilberry extract (200 mg/kg body weight [BW] dissolved in 1 mL of saline) daily for 10 consecutive days. CP-treated animals were administered a single intraperitoneal dose (7.5 mg/kg BW). Finally, a bilberry+CP group received oral bilberry extract (200 mg/kg BW) daily for 10 consecutive days, with one intraperitoneal dose of CP (7.5 mg/kg BW) on day 2. We assessed sperm count, motility, viability, and abnormalities, along with testis weight, testis weight-to-BW ratio, antioxidant activity, levels of oxidative stress markers (malondialdehyde [MDA] and hydrogen peroxide [H2O2]), sex hormones (follicle-stimulating hormone [FSH], luteinizing hormone [LH], and testosterone), and apoptotic and anti-apoptotic markers, and DNA damage. Testicular tissue underwent histopathological examination. Among CP-treated rats, significantly lower values were observed for testis weight; testis weight-to-BW ratio; levels of FSH, LH, testosterone, superoxide dismutase, catalase, glutathione S-transferase, glutathione, and B-cell lymphoma 2; and sperm count, motility, and proportion of normal sperm. CP administration was associated with higher MDA, H2O2, p53, Bax, cytochrome c, caspase 9, and caspase 3 levels, along with elevated tail moment. However, bilberry extract administration significantly improved all altered parameters. Bilberry treatment demonstrated protective effects and reduced CP-induced testicular toxicity via antioxidant activity and cytoprotection.

The Antioxidant and Anti-Inflammatory Properties of Wild Bilberry Fruit Extracts Embedded in Mesoporous Silica-Type Supports: A Stability Study.

Polyphenolic extracts from wild bilberries (Vaccinium myrtillus L.) have shown antioxidant and anti-inflammatory effects, but they are prone to degradation when exposed to environmental factors, limiting their use in biomedical applications. To overcome this issue, this study proposed the embedding of wild bilberry fruit ethanolic extracts in pristine mesoporous silica functionalized with organic groups (mercaptopropyl and propionic acid), as well as coated with fucoidan, a biopolymer. Herein, we report a stability study of free and incorporated extracts in mesoporous silica-type supports in high-humidity atmospheres at 40 °C up to 28 days, using HPLC analysis, thermal analysis, and radical scavenging activity determination. Better chemical and thermal stability over time was observed when the extracts were incorporated in mesoporous silica-type supports. After 12 months of storage, higher values of antioxidant activity were determined for the extract embedded in the supports, silica modified with mercaptopropyl groups (MCM-SH), and fucoidan-coated silica (MCM-SH-Fuc) than that of the free extract due to a synergistic activity between the support and extract. All encapsulated extracts demonstrated remarkable effects in reducing NO production in LPS-stimulated RAW 264.7 cells. The treatment with extract embedded in MCM-SH-Fuc in a dose of 10 μg/mL surpassed the effect of free extract in the same concentration. For the extract encapsulated in an MCM-SH support, a lower IC50 value (0.69 μg/mL) towards COX-2 was obtained, comparable with that of Indomethacin (0.6 μg/mL). Also, this sample showed a higher selectivity index (2.71) for COX-2 than the reference anti-inflammatory drug (0.98). The developed formulations with antioxidant and anti-inflammatory properties could be further used in nutraceuticals.

Anti-Aging Properties of Chitosan-Based Hydrogels Rich in Bilberry Fruit Extract.

Photoaging is a process related to an increased level of reactive oxygen species (ROS). Polyphenols can scavenge free radicals in the body, which can delay skin aging. Therefore, our work aimed to prepare a biologically active extract from dry fruits of Vaccinium myrtillus or Vaccinium corymbosum and use it for the preparation of hydrogels for topical application. Therefore, eight different extracts (using V. myrtillus and V. corymbosum and different extraction mixtures: methanol, methanol-water 1:1, water, acetone-water 1:1) were prepared and their phytochemical (total polyphenolic content, total flavonoid content, total anthocyanin content) and biological properties (antioxidant, anti-hyaluronidase, and anti-tyrosinase activity) were assessed. Cytotoxicity towards HaCaT keratinocytes was also determined. Based on the results, the acetone-water extract from V. myrtillus was selected for further study. Using the Design of Experiments approach, chitosan-based hydrogels with bilberry fruit extract were prepared. The content of extract and chitosan were selected as independent factors. The activity of hydrogels depended on the extract content; however, the enzyme-inhibiting (anti-hyaluronidase and anti-tyrosinase) activity resulted from the presence of both the extract and chitosan. Increased concentration of chitosan in the hydrogel base led to increased viscosity of the hydrogel and, consequently, a slower release of active compounds. To get optimal hydrogel characteristics, 1% extract and 2.5% MMW chitosan were utilized. The research suggests the validity of using bilberry fruit extracts in topical preparations with anti-aging properties.

Anthocyanins as an element of nutritional support for athletes: effects and molecular mechanisms of action

Antioxidants are among the common components of sports nutrition designed to counteract oxidative stress that develops during intense physical activity. One of the promising antioxidants are anthocyanins which belong to polyphenolic compounds of plant origin (class of flavonoids). The purpose of the research was to analyze the results of prospective controlled studies on the effect of anthocyanins on physical performance, and to consider the possible molecular mechanisms of their action. Material and methods. Sources were searched in PubMed, Google Scholar, and CyberLeninka databases of peer-reviewed scientific literature without restrictions on the year of publication using the following keywords: anthocyanins, physical performance, recovery, sport and exercise nutrition, oxidative stress, inflammation. Results. The main data set on the effects of anthocyanins in athletes was obtained using extracts of blackcurrant and Montmorency tart cherry. Volunteers received anthocyanins at a dose of 86-547 mg per day for 1 to 10 days with subsequent evaluation of their performance in cycling, running and fitness activities. The possibility of favorable effect of anthocyanins on physical performance and acceleration of its recovery after exertion has been shown. The source, dose and duration of intake did not significantly influence the established effects of anthocyanins. Acting as exogenous regulators of metabolism, anthocyanins can activate several mechanisms of performance enhancement, including influence on antioxidant and immune status and apoptosis intensity. Anthocyanins prevent the formation of reactive oxygen species, neutralize electrophilic compounds by direct interaction or through activation of Nrf2 factor, which regulates the transcription of antioxidant enzyme genes. The basis of the anti-inflammatory action of anthocyanins is their ability to inhibit MARK and NF-κB mediated signal transduction. Inclusion of bilberry and blackcurrant extract in the diet prevented the intensification of myocyte apoptosis and suppression of cellular immunity induced by exhausting exercise. An additional mechanism of anthocyanin action on physical performance may be an increase in blood supply of organs and tissues due to vascular dilation caused by activation of endothelial nitric oxide synthase. Conclusion. The intake of plant extracts with a high anthocyanin content can increase physical performance and improve recovery after physical exertion, which may be due to the antioxidant and anti-inflammatory effects of anthocyanins, their ability to regulate apoptosis processes and improve blood supply to organs and tissues.

Exploring the potential of bilberry extracts as natural antifungal and bioherbicidal agents in agriculture: Composition and antioxidant activity

This study aims to investigate the composition of the leaf essential oil and the total phenolic and flavonoid contents as well as the antioxidant activity of the n-hexane, ethyl acetate, and methanol extracts of Vaccinium myrtillus leaves collected from two different locations (Mu?la and Mersin, Turkey). In addition, the antifungal activity against Sclerotinia sclerotiorum (SS), Fusarium oxysporum f sp. melonis (FOM), Fusarium oxysporum f sp. cucumerinum (FOC), and Rhizoctonia solani (RS) and the bioherbicidal activity against Taraxacum officinale and Rumex crispus were evaluated for the first time. The methanolic extract inhibited mycelium growth of SS, FOM, FOC, and RS in a dose-dependent manner. No significant difference was observed in antifungal activity between the two different collection sites. In the bioherbicidal activity tests, the methanol extract of the fruits completely suppressed the root-shoot development of Taraxacum officinale, while also significantly inhibiting the root-shoot development during the seed germination of Rumex crispus compared to the control group at 3 mg/mL concentration for both locations. Based on the results of the GC/MS analysis, the major constituents identified in the leaf essential oils collected from the Mu?la and Mersin locations were ?-pinene (29.16%/15.75%), eucalyptol (22.19%/26.46%), linalool (12.66%/25.27%), and linalyl acetate (7.43%/2.71%). The total phenolic and flavonoid contents of the plant extracts for the Mu?la and Mersin samples were determined as (42.80-157.58 GAE/g extracts), (29.38-151.44 GAE/g extracts) and (10.52-37.88 QE/g extracts), (14.81-44.18 QE/g extracts) respectively. In addition, the plant exhibited significant antioxidant activity. These findings indicate quantitative differences in the chemical composition of the essential oils between the two geographical locations. These findings are significant as they provide insights for the development of new natural antifungal agents with potential applications in agriculture.

Investigation of the Effects of Bilberry Extract on Methotrexate-Induced Hepatotoxicity

Amaç: Metotreksat (Mtx), kanser ve otoimmün hastalıkların tedavisinde yaygın olarak kullanılmaktadır. Ancak Mtx, kendisi gibi birçok kemoterapötiğin metabolizmasından sorumlu olan karaciğerde toksisiteye neden olabilir. Yaban mersini (Ym) meyvesi, yapısında yoğun olarak bulunan fenolik bileşikler sayesinde antioksidan ve antienflamatuvar etkiye sahiptir. Bu çalışma, Ym ekstresinin, Mtx kaynaklı hepatotoksisiteye olan etkilerini araştırmayı amaçlamakta-dır. Materyal ve metod: Çalışma için 21 adet Spraque Dawley ırkı erişkin sıçan, sayıca eşit üç gruba bölündü. Bir gruba, deneyin ilk günü tek doz 30 mg/kg intraperitoneal (ip.) Mtx verildi (MtxG). Diğer gruba tek doz 30 mg/kg ip. Mtx ve beş gün boyunca günde bir kez oral gavajla 200 mg/kg Ym ekstresi verildi (Mtx+YmG). Kontrol grubuna ise herhangi bir işlem yapılmadı (KG). Deney sonunda alınan karaciğer dokularının bir kısmı histolojik olarak değer-lendirilerek skorlanırken diğer kısmı da biyokimyasal olarak değerlendirildi. Bulgular: MtxG’nin karaciğer dokularında malondialdehid (MDA) ve kaspaz 3 düzeyleri KG’ye göre yüksekti. KG’ye göre anlamlı olmamakla birlikte MtxG’de, 8-hidroksi-2′-deoksiguanozin (8-OHdG) ve oksidatif stres indeksi (OSİ)’nde artış; süperoksit dismutaz (SOD), katalaz (CAT) ve total antioksidan kapasite (TOS)’de ise azalma mevcut-tu. Mtx+YmG’de MDA düzeyleri MtxG’ye göre azaldı. Bu grupta, kaspaz 3, OSI ve 8-OHdG düzeylerinde de MtxG'ye göre istatistiksel olarak anlamlı olmayan belirgin bir azalma görüldü. Ayrıca bu gruptaki SOD seviyesi KG’ye göre düşükken CAT ve total antioksidan kapasite (TAS) seviyeleri ise MtxG’ye göre istatistiksel anlamlı olmasa da yüksekti. MtxG’de; total hasar skorunda, hepatositlerdeki vakuolizasyon/dejenerasyon/piknoziste, Kupffer hücrelerinde ve vazodilatasyon/konjesyonda kontrole göre artış mevcuttu. Mtx+YmG’de ise tüm bu histopatolojik bulgularda ve total hasar skorunda azalma mevcuttu. Sonuç: Oral yolla uygulanan Ym ekstresi, sıçan karaciğerindeki Mtx kaynaklı oksidatif stresi, apoptozu ve bunların neden olduğu hepatotoksisiteyi azaltabilir.

THE AMELIORATING EFFECTS OF BILBERRY ON METHOTREXATE-INDUCED INTESTINAL INJURY

Objective: Methotrexate (MTX) is a folic acid analogue chemotherapeutic used in the treatment of some malignant tumors and autoimmune diseases. In addition to its antitumoral properties, it can also exhibit toxic effects on the kidney and intestines. Bilberry (BB) fruit is a potent natural antioxidant rich in anthocyanidins. The purpose of this study was to investigate the potential ameliorating effects of BB against MTX-induced intestinal damage using biochemical and histological methods.
 Material and Methods: Twenty-one adult female Sprague Dawley rats were divided into three equal groups. No procedure was performed on the control group (ControlG), while the MTX group (MTXG) and MTX+BB extract group (MTX+BBG) received a single intraperitoneal dose of 30 mg/kg MTX on the first day of the experiment. MTX+BBG also received 200 mg/kg BB extract by oral gavage once daily for five days starting on the first day of the experiment. Half the intestinal tissues removed from the duodenal region at the experiment were used for biochemical evaluation, and the other half for histological examination.
 Results: Malondialdehyde (MDA), total oxidant status (TOS), oxidative stress index (OSI), and 8-oxo-2′-deoxyguanosine (8- OHdG) values were all higher in MTXG intestinal tissues than in ControlG. MDA, TOS, OSI, and 8-OHdG values were lower in MTX+BBG than in MTXG. In addition, caspase-3 levels in MTXG were higher than those in both ControlG and MTX+BBG, while total antioxidant status (TAS) values were lower. In terms of histology, villous hemorrhage, inflammatory cell infiltration, fusion, and degeneration in the villus epithelium were present in MTXG intestinal tissue, and the total damage score was also high. Improvement in all these parameters was present in MTX+BBG.
 Conclusion: Orally administered BB extract can improve MTX-induced intestinal damage through its antioxidant and anti-inflammatory effects.

PH-responsive color-indicating film of pea protein isolate cross-linked with dialdehyde carboxylated cellulose nanofibers for pork freshness monitoring

The limited mechanical performance and responsiveness of protein-based smart packaging materials have hindered their development. To address these issues, this study prepared a pH-responsive smart film by introducing dialdehyde carboxylated cellulose nanofibers (DCCNFs) as the cross-linking agent capable of covalently reacting with proteins, and bilberry extract (BE) as a pH-responsive indicator into pea protein isolate (PPI) matrix. The results demonstrated that adding DCCNF and BE enhanced the PPI film's thermal stability, density, and UV barrier properties. Tensile tests revealed significant improvements in both tensile strength and elongation at the break for the resulting film. Furthermore, films containing DCCNF and BE exhibited lower moisture content, swelling ratio, water vapor permeability, and relative oxygen transmission compared to PPI films. Notably, the anthocyanins in BE endowed the film with visual color changes corresponding to different pH values. This feature enabled the film to monitor pork freshness; a transition from acidic to alkaline in pork samples was accompanied by a color change from brown to brownish green in the film as storage time increased. Overall, these findings highlight that this developed film possesses excellent physicochemical properties and sensitive pH response capabilities, making it a promising candidate for future smart packaging applications.

Potential Chemopreventive Effects of Dietary Combination of Phytochemicals against Cancer Development.

Cancer remains a major cause of cancer-related death worldwide. Over 70% of epithelial malignancies are sporadic and are related to lifestyle. Epidemiological studies suggest an inverse correlation between cancer incidence and fruit and vegetable intake. Numerous preclinical studies using in vitro (cell lines) and in vivo animal models of oncogenesis have reported the chemopreventive effects of dietary phytochemical agents through alterations in different biomarkers and signaling pathways. However, there is contrasting evidence from preclinical studies and clinical trials. To date, the most studied compounds include curcumin, resveratrol, isoflavones, green tea extract (epigallocatechin gallate), black raspberry powder (anthocyanins and ellagitannins), bilberry extract (anthocyanins), ginger extract (gingerol derivatives), and pomegranate extract (ellagitannins and ellagic acid). Overall, the clinical evidence of the preventive effects of dietary phytochemicals against cancer development is still weak, and the amount of these phytochemicals needed to exert chemopreventive effects largely exceeds the common dietary doses. Therefore, we propose a combination treatment of natural compounds that are used clinically for another purpose in order to obtain excess inhibitory efficacy via low-dose administration and discuss the possible reasons behind the gap between preclinical research and clinical trials.

Src Tyrosine Kinase Inhibitory and Antioxidant Activity of Black Chokeberry and Bilberry Fruit Extracts Rich in Chlorogenic Acid.

Edible berries such as the fruits of black chokeberry (Aronia melanocarpa (Michx.) Elliott) and bilberry (Vaccinium myrtillus L.) are considered to be rich in phenolic compounds, which are nowadays attracting great interest due to their promising health benefits. The main objective of our study was to investigate, for the first time, their inhibitory properties on Src tyrosine kinase activity, as this enzyme plays an important role in multiple cellular processes and is activated in both cancer and inflammatory cells. In hydroethanolic fruit extracts, 5.0-5.9% of total polyphenols were determined spectrophotometrically, including high amounts of hydroxycinnamic acid derivatives. HPLC analysis revealed that the black chokeberry and bilberry extracts contained 2.05 mg/g and 2.54 mg/g of chlorogenic acid, respectively. Using a time-resolved fluorescence resonance energy transfer (TR-FRET) assay, the extracts studied were found to have comparable inhibitory effects on Src tyrosine kinase, with IC50 values of 366 µg/mL and 369 µg/mL, respectively. The results also indicated that chlorogenic acid contributes significantly to the observed effect. In addition, both fruit extracts exhibited antioxidant activity by scavenging DPPH and NO radicals with SC50 values of 153-352 µg/mL. Our study suggested that black chokeberry and bilberry fruits may be beneficial in cancer and other inflammation-related diseases.

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.

735-P: Baseline Characteristics in the REVERSE-IT Study, a 24-Week, 600-Person Randomized Placebo-Controlled Study of the Metabolic Effects of the Combination of Five Plant Extracts TOTUM-63 (T63)

The role of natural based products to support glucose control in people with prediabetes or early type 2 diabetes (T2D) (i.e. dysglycemia) is debated. T63 is a polyphenol-rich substance composed by artichoke, chrysanthellum, olive leaf, bilberry, and black pepper extracts, and studied in REVERSE-IT (NCT04423302), an ongoing 24 weeks, multi-center, multi-country clinical trial involving people with dysglycemia. It tests the hypothesis that 5 g T63 vs placebo (PBO) leads to significant reduction in fasting plasma glucose, while secondary outcomes include effects on lipids, blood pressure (BP), weight and waist circumference. The confirmatory primary analysis will compare T63 vs PBO when provided TID (double-blinded) and is powered based on minimum 174 completers in each arm. Effects of T63 BID vs PBO will be analyzed open label. A total of n=627 were recruited over 2 years from 61 sites across 7 European countries. Mean age is 57 years, BMI 32.2 kg/m2, and HbA1c 6.1% (table 1). The majority have prediabetes (78.3%), with a better glycemic profile and BP compared to those with T2D (21.7%). REVERSE-IT has enrolled an appropriate population to help inform clinical decision making on the use of plant-extract based T63 to support glucose homeostasis in people with dysglycemia. Readout is expected in the second half of 2023. Disclosure Y.F.Otero: Employee; Valbiotis. T.Maugard: None. M.Cazaubiel: Board Member; Valbiotis. B.Pereira: None. B.Guigas: Advisory Panel; Valbiotis, Research Support; Valbiotis. S.Hadjadj: Consultant; Bayer Inc., Novo Nordisk, Valbiotis, Research Support; AstraZeneca, Novartis, Speaker's Bureau; Abbott Diagnostics, Boehringer-Ingelheim. S.Peltier: Board Member; Valbiotis. A.Marette: Advisory Panel; Valbiotis, Amancia, Plexus, Acasti. J.Bard: Consultant; Valbiotis, Biofortis France, Stock/Shareholder; Valbiotis. P.Sirvent: Board Member; Valbiotis. O.Johansen: Employee; Nestlé Health Science. V.Chavanelle: Employee; Valbiotis. M.Bargetto: Employee; Valbiotis. V.Sapone: None. A.Bouchard-mercier: Employee; Valbiotis. F.Le joubioux: Employee; Valbiotis. N.Boisseau: None.

A grape seed and bilberry extract reduces blood pressure in individuals at risk of developing type 2 diabetes: the PRECISE study, a double-blind placebo-controlled cross-over intervention study.

Type 2 Diabetes Mellitus (T2DM) is a major risk factor for the development of cardiometabolic diseases. T2DM prevention is largely based on weight-loss and whole diet changes, but intervention with dietary plant bioactives may also improve metabolic health. To assess whether supplementation with bilberry and grape seed extract for 12 weeks improves cardiometabolic outcomes in individuals at risk of developing T2DM, and to determine whether individual treatment response is associated with differences in gut microbiota composition and levels of phenolic metabolites in blood and feces. In the randomized, double-blind, placebo-controlled, cross-over PRECISE intervention study, 14 participants, aged ≥45 years, with a BMI >28 kg/m2, and having an increased risk of T2DM, received a supplement containing 250 mg of bilberry plus 300 mg of grape seed extract, or 550 mg of a control extract, per day, for 12 weeks each. Blood samples were obtained for the assessment of HbA1c, fasting glucose, oral glucose tolerance tests, insulin, glucagon levels, total, LDL and HDL cholesterol, and phenolic acids. We also assessed advanced glycation end products in the skin, ambulatory 24 hours blood pressure, 7-day dietary intake by weighed food diaries, fecal levels of phenolic metabolites using LC-MS/MS and gut microbiota composition using 16S rRNA gene sequencing analysis. The combined bilberry and grape seed extract did not affect glucose and cholesterol outcomes, but it decreased systolic and diastolic ambulatory blood pressure by 4.7 (p < 0.001) and 2.3 (p = 0.0009) mmHg, respectively. Eight out of fourteen participants were identified as blood pressure 'responders'. These responders had higher levels of phenylpropionic and phenyllactic acids in their fecal samples, and a higher proportional abundance of Fusicatenibacter-related bacteria (p < 0.01) in their baseline stool samples. Long-term supplementation with bilberry and grape seed extract can improve systolic and diastolic blood pressure in individuals at risk of T2DM. Individual responsiveness was correlated with the presence of certain fecal bacterial strains, and an ability to metabolize (epi)catechin into smaller phenolic metabolites.Clinical trial registry number: Research Registry (number 4084).

Bilberry-containing supplements on severe dry eye disease in young and middle-aged adults: A 3-month pilot analysis.

To explore the effect of bilberry and fish oil combination supplement on a small clinical sample patient-base with severe dry eyes. Twenty-four subjects were recruited with twelve randomly assigned to the intervention and control groups, respectively. Inclusion criteria included severe dry eye symptoms determined by scores >33 from the Ocular Surface Disease Index (OSDI) questionnaire. The intervention group was instructed to take an oral supplement with key ingredients of 600 mg bilberry extract and 240 mg docosahexaenoic acid-refined fish oil once daily for 3 months. The control group did not take any supplements. Mean changes in OSDI score, non-invasive tear break-up time (NITBUT), phenol red thread test (PRT), and percentage of meibomian gland openings were used as outcome measures. Testing was done at baseline, 1-month, and 3-month follow-up. Comparison between the treatment and control groups, and the younger adult and middle-age groups were performed. The mean baseline values for the treatment and control groups were not clinically different. The OSDI score, NITBUT, PRT, and percentage of meibomian gland openings improved after taking the supplements for 3 months. The OSDI score, NITBUT, and PRT showed clinical improvements between the intervention and control groups. These improvements were consistent between the two age groups. This study suggested preliminary improvements in signs and symptoms of severe dry eyes that were independent of age after taking dietary supplementation of bilberry extract and fish oil for 3 months. Further studies using more device-based measures and a placebo supplement are warranted.