Prussian blue nanocubes with peroxidase-like activity for polyphenol detection in commercial beverages.

The present study describes an efficient method for the determination of polyphenol content in beverages based on a composite material of graphene oxide decorated with Prussian blue nanocubes (rGO/PBNCs). In this method, rGO/PBNCs act as a nanoenzyme with peroxidase-like catalytic activity and produce a colorimetric product in the presence of hydrogen peroxide and tetramethylbenzidine (TMB). To verify the effectiveness of the method, we used two model standards for antioxidants: gallic acid (GA) and tannic acid (TA). The method validation included a comparison of the performance of a natural enzyme and an artificial one (rGO/PBNCs) and two polyphenols in the analysis of commercial beverage samples. After optimization, a pH of 4, ambient temperature (22 °C), a reaction time of 2 minutes and an rGO/PBNCs concentration of 0.01 µg mL-1 were found to be the most favorable conditions. The detection limits obtained were 5.6 µmol L-1 for GA and 1.5 µmol L-1 for TA. Overall, rGO/PBNCs offer advantages over natural enzymes in terms of stability, versatility, scalability and durability, making them attractive candidates for a wide range of catalytic and sensory applications.

Nanozymes sensor array for discrimination and intelligent sensing of phenolic acids in food.

Although nanozymes sensor arrays have the potential to recognize multiple target substances simultaneously, they currently rarely identify phenolic acids in food due to limited catalytic performance and complex preparation conditions of nanozymes. Here, inspired by the structure of polyphenol oxidase, we have successfully prepared a novel gallic acid-Cu (GA-Cu) nanozyme with laccase-like activity. Due to the different catalytic efficiency of GA-Cu nanozymes towards six common phenolic acids, a three-channel colorimetric sensor array was constructed using reaction kinetics as the sensing unit to achieve high-throughput detection and identification of six phenolic acids within a concentration range from 1 to 100 µM. This method avoids the creation of numerous sensing units. Notably, the successful discrimination of six phenolic acids in samples of juice, beer, and wine has been achieved by the sensor array. Finally, aided by smartphones, a portable technique has been devised for the detection of phenolic acids.

Effects of post-fermentation addition of green tea extract for sulfur dioxide replacement on Sauvignon Blanc wine phenolic composition, antioxidant capacity, colour, and mouthfeel attributes.

This study examines the feasibility of replacing SO2 in a New Zealand Sauvignon Blanc wine with a green tea extract. The treatments included the control with no preservatives (C), the addition of green tea extract at 0.1 and 0.2 g/L (T1 and T2), and an SO2 treatment at 50 mg/L (T3). Five monomeric phenolic compounds were detected in the green tea extract used for the experiment, and their concentrations ranged in the order (-)-epigallocatechin gallate > (-)-epigallocatechin > (-)-epicatechin > (-)-epicatechin gallate > gallic acid. At the studied addition rates, these green tea-derived phenolic compounds contributed to ∼70% of the antioxidant capacity (ABTS), ∼71% of the total phenolic index (TPI), and âˆ¼ 84% of tannin concentration (MCPT) of the extract dissolved in a model wine solution. Among wine treatments, T1 and T2 significantly increased the wine's colour absorbance at 420 nm, MCPT, gallic acid and total monomeric phenolic content. TPI and ABTS were significantly higher in wines with preservatives (i.e., T2 > T1 â‰… T3 > C, p < 0.05). These variations were observed both two weeks after the treatments and again after five months of wine aging. Additionally, an accelerated browning test and a quantitative sensory analysis of wine colour and mouthfeel attributes were performed after 5 months of wine aging. When exposed to excessive oxygen and high temperature (50 °C), T1 and T2 exhibited ∼29% and 24% higher browning capacity than the control, whereas T3 reduced the wine's browning capacity by ∼20%. Nonetheless, the results from sensory analysis did not show significant variations between the treatments. Thus, using green tea extract to replace SO2 at wine bottling appears to be a viable option, without inducing a negative impact on the perceptible colour and mouthfeel attributes of Sauvignon Blanc wine.

Chemical composition and potential antioxidant, anti-inflammatory, and analgesic efficacy of Cistus albidus L.

This study aims to assess the chemical composition of the aqueous extract of Cistus albidus L. leaves, as well as the potential of aqueous and hydroethanol extracts of the leaves and seeds as analgesic, anti--inflammatory, and antioxidant agents. The contents of phenolics and inorganic constituents were determined in C. albidus seeds and leaves; antioxidant capacity was assessed by 3 complementary and diverse tests. The carrageenan-induced paw edema technique was used to investigate the anti-inflammatory effect in vivo, and albumin denaturation to evaluate the anti-inflammatory effect in vitro. The acetic acid-induced contortion test, the tail-flick test, and the plantar test were used to assess the analgesic effi cacy in vivo. Chemical analysis was performed by UPLC-MS/MS to quantify several phenolic compounds including catechin (1,627.6 mg kg-1), quercitrin (1,235.8 mg kg-1) and gallic acid (628. 2 mg kg-1). The ICP analysis revealed that potassium and calcium were the main inorganic components in the seeds and leaves of C. albidus. The hydroethanolic extract of the leaves showed the highest content of polyphenols/flavonoids, whereas the highest value of proantho cyanidins was detected in the aqueous extract of the seeds. All extracts showed potent antioxidant activity related to different phenolic compounds (quercetin, gallic acid, astragalin, catechin, and rutin). The aqueous extract of the leaves strongly inhibited paw edema (76.1 %) after 6 h of treatment and showed maximal inhibition of protein denaturation (191.0 µg mL-1 for 50 % inhibition) and analgesic activity in different nociceptive models. The presented data reveal that C. albidus extracts potentially show antioxidant, anti-inflammatory, and analgesic activities that could confirm the traditional use of this plant.

Unraveling the potential nutritional benefits of fermented date syrup waste: Untargeted metabolomics and carbohydrate metabolites of in vitro digested fraction.

Valorization of fruit by-products is a crucial area of research for the development of innovative bio-based products. This study investigated the physicochemical properties and health-promoting benefits of date syrup waste, both fermented by Pichia cecembensis or Pichia kudriavzevii (FDSW), and unfermented (CDSW). Metabolomics profiles of these samples were identified post in vitro digestion. FDSW exhibited 42 volatile compounds, including 9 new ones, and contained (-)-epicatechin, tyrosol, and gallic acid. Bioaccessible fractions of FDSW demonstrated substantial α-amylase inhibition, with percentages of 40.7 % and 53.9 %, respectively. FDSW displayed superior cytotoxicity against Caco2 and MCF-7 cancer cell lines, with an average of âˆ¼75 % and 56 %, respectively. Untargeted metabolomics analysis revealed an increase in secondary metabolites, totaling 27 metabolites. LC-QTOF analysis of bioaccessible carbohydrate metabolites in FDSW identified two phytochemical groups, alkaloids, and terpenoids. This study underscores the potential of FDSW for producing value-added bio-based products with desirable characteristics and health benefits.

Exhausted Grape Seed Residues as a Valuable Source of Antioxidant Molecules for the Formulation of Biocompatible Cosmetic Scrubs.

Grape seed of Obeidi, a white Lebanese autochthonous variety, was previously tested in different studies as a valuable source of bioactive molecules such as polyphenols, oils, and proteins by means of extraction procedures for the development of cosmetic and therapeutic products. However, an un-valorized, exhausted grape seed residue remains as "secondary waste" after the extraction processes. In this study, the exhausted seeds have been further exploited to produce cosmetic scrubs capable of releasing antioxidant molecules during the exfoliation process, in accordance with the principles of the circular economy and going toward a zero-waste process. The deep characterization of the exhausted seeds confirmed the presence of antioxidant phenolic molecules including gallic acid, catechins and protocatechuic acid (0.13, 0.126, and 0.089 mg/g of dry matter DM), and a high phenolic content (11.85 mg gallic acid equivalents (GAE)/g of dry matter (DM)). Moreover, these residues were shown to possess a sandy texture (Hausner ratio (HR): 1.154, Carr index (CI): 0.133, and angle of repose: 31.62 (°) degrees), similar to commercial natural exfoliants. In this respect, exhausted Obeidi grape seed residues were incorporated at increasing concentrations (0.5, 1, 1.5, and 2% w/w) in a cosmetic scrub, and stored for 5 weeks at 4, 25, and 50 °C for stability testing. All tested scrub formulations exhibited good spreadability with a spread diameter of 3.6-4.7 cm and excellent physical stability, as no phase separation or color change were observed after four cycles of heat shock at 4 and 50 °C. Finally, an in vivo skin irritation test showed that the scrub enriched with 1.5% of exhausted Obeidi grape seed residues was the most promising formulation, as it possessed a high amount of phenolic molecules (0.042 ± 0.001 mg GAE/mL of scrub) and good stability and could be safely applied to the skin with no irritation phenomena. Overall results underlined that exhausted grape seed residues can be transformed into promising systems for both physical and chemical exfoliation, thus confirming the importance of the effective exploitation of agro-industrial by-products for the development of high value cosmeceutics towards a more sustainable and zero-waste approach.

Isolation and characterization of natural inhibitors of post-proline specific peptidases from the leaves of Cotinus coggygria Scop.

ETHNOPHARMACOLOGICAL RELEVANCE: Cotinus coggygria has a number of applications in traditional medicine most of which are connected with its anti-inflammatory and anti-oxidant properties. Since inflammation and oxidative stress are recognized as triggering factors for cancer, anti-cancer activity has also been documented and the possible mechanisms of this activity are under investigation. Important components of C. coggygria extracts are shown to be hydrolysable gallotannins of which pentagalloyl-O-glucose has been studied in details. This compound inhibits various enzymes including prolyl oligopeptidase which is involved in tumorigenesis and tumour growth. According to our pilot studies, oligo-O-galloylglucoses with more than five galloyl residues are also presented in the herb of Bulgarian origin, but their activities have not been examined. AIM OF THE STUDY: To establish an extraction method by which it is possible to concentrate high molecular hydrolysable gallotannins from dried leaves of Cotinus coggygria and to determine their inhibitory properties towards prolyl oligopeptidase and fibroblast activation protein α. MATERIALS AND METHODS: Dried leaves of C. coggygria were extracted using different solvents in single-phase or biphasic systems under various extraction conditions. Main compounds of the extracts were identified by using high performance liquid chromatography and liquid chromatography - high resolution mass spectrometry. The extracts' inhibitory properties towards prolyl oligopeptidase and fibroblast activation protein α were studied on recombinant human enzymes by enzyme kinetic analyses using a fluorogenic substrate. RESULTS: Ethyl acetate/water (pH 3.0) extraction of dried plant leaves proved to be the most efficient method for isolation of high molecular hydrolysable gallotannins which can be further concentrated by precipitation of dicyclohexylammonium salts in ethyl acetate. The main components of those extracts were oligo-O-galloyl glucoses with more than five gallic acid residues. They were shown to inhibit both enzymes studied but were about 30 times more effective inhibitors of prolyl oligopeptidase. CONCLUSIONS: C. coggygria from Bulgarian origin is shown to possess a substantial quantity of oligo-O-galloyl glucoses with more than five gallic acid residues which has not been described thus far in the same herb from other sources. An extraction method useable for concentrating those compounds is established. They are found to inhibit prolyl oligopeptidase with a very good selectivity to fibroblast activation protein α. The previously described antitumor activity of this plant may be at least in part due to the inhibition of the above enzymes which has been shown to participate in the genesis and development of various types of tumors.

Chemical Profiling and Therapeutic Evaluation of Standardized Hydroalcoholic Extracts of Terminalia chebula Fruits Collected from Different Locations in Manipur against Colorectal Cancer.

Terminalia chebula Retz. (Fam. Combretaceae), locally called Manahei, is a well-known medicinal plant that grows wildly in Manipur, a Northeastern state of India. It is used as a mild laxative, an anti-inflammatory agent, and a remedy for piles, colds, and ulcers by ethnic communities of the state. The hydroalcoholic extract obtained from four fruit samples of T. chebula collected from different locations in Manipur were analyzed using gas chromatography-mass spectrometry (GC-MS) and high-performance thin-layer chromatography (HPTLC) for their chemical constituents and evaluated for their anticancer activity against the colon cancer cell HCT 116. GC-MS analysis results indicated significant variation in the composition and percentage of major compounds present in the extracts. 1,2,3-Benzenetriol was the most abundant chemical constituent present in all four extracts of T. chebula, ranging from 20.95 to 43.56%. 2-Cyclopenten-1-one, 5-hydroxymethylfurfural, and catechol were commonly present in all extracts. Two marker compounds, gallic acid and ellagic acid, were also quantified usingHPTLC in all four extracts of T. chebula. The highest content of gallic acid (22.44 ± 0.056 µg/mg of dried extract) was observed in TCH, and that of ellagic acidwas found in TYH (11.265 ± 0.089 µg/mg of dried extract). The IC50 value of TYH for the DPPH and ABTS assays (12.16 ± 0.42 and 7.80 ± 0.23 µg/mL) was found to be even lower than that of Trolox (18 ± 0.44 and 10.15 ± 0.24 µg/mL), indicating its strong antioxidant properties among the four extracts of T. chebula. The MTT assay determined the effect of T. chebula extracts on the viability of HCT 116 cells. TYH showed the highest activity with anIC50 value of 52.42 ± 0.87 µg/mL, while the lowest activity was observed in TCH (172.05 ± 2.0 µg/mL). The LDH assay confirmed the cytotoxic effect of TYH in HCT 116 cells. TYH was also found to induce caspase-dependent apoptosis in HCT 116 cells after 48 h of treatment. Our study provides insight into the diversity of T. chebula in Manipur and its potential activity against colon cancer.

Bioactive Compounds, Antioxidant Activities, and HPLC Analysis of Nine Edible Sprouts in Cambodia.

The non-nutritional health benefits of sprouts are unconfirmed. Thus, nine sprout methanolic extracts were tested for phytoconstituents and antioxidant activity. The TPC, TCC, TFC, TAC, and TALC were measured. ABTS and DPPH radical scavenging and ferric-reducing antioxidant power assays were used to assess the antioxidant activity. HPLC detected gallic acid, vanillin, syringic acid, chlorogenic acid, caffeic acid, and rutin in the extracts. The sprout extracts contained six compounds, with caffeic acid being the most abundant. Gallic acid, syringic acid, chlorogenic acid, caffeic acid, vanillin, and rutin were highest in soybean, black sesame, mustard, sunflower, white radish, and black sesame sprouts, respectively. Sunflower sprouts had the highest level of TCC while soybean sprouts had the highest level of TFC, Taiwanese morning glory had the highest level of TPC, mustard sprouts had the highest level of TALC, and black sesame sprouts had the highest level of TAC. Taiwanese morning glories scavenged the most DPPH and ABTS radicals. Colored and white radish sprouts had similar ferric-reducing antioxidant power. Antioxidation mechanisms varied by compound. Our findings demonstrated that sprouts have biological effects, and their short time for mass production offers an alternative food source for health benefits, and that they are useful for future research development of natural products and dietary supplements.

Electrochemical Sensing of Gallic Acid in Beverages Using a 3D Bio-Nanocomposite Based on Carbon Nanotubes/Spongin-Atacamite.

Gallic acid (GA) is one of the most important polyphenols, being widely used in the food, cosmetic, and pharmaceutical industries due to its biological effects such as antioxidant, antibacterial, anticancer, antiviral, anti-inflammatory, and cardioprotective properties. Hence, simple, fast, and sensitive determination of GA is of particular importance. Considering the fact that GA is an electroactive compound, electrochemical sensors offer great potential for GA quantitation due to their fast response time, high sensitivity, and ease of use. A simple, fast, and sensitive GA sensor was fabricated on the basis of a high-performance bio-nanocomposite using spongin as a natural 3D polymer, atacamite, and multi-walled carbon nanotubes (MWCNTs). The developed sensor showed an excellent response toward GA oxidation with remarkable electrochemical features due to the synergistic effects of 3D porous spongin and MWCNTs, which provide a large surface area and enhance the electrocatalytic activity of atacamite. At optimal conditions by differential pulse voltammetry (DPV), a good linear relationship was obtained between peak currents and GA concentrations in a wild linear range of 500 nM to 1 mM. Subsequently, the proposed sensor was used to detect GA in red wine as well as in green and black tea, confirming its great potential as a reliable alternative to conventional methods for GA determination.

An integrated approach to the structural characterization, long-term toxicological and anti-inflammatory evaluation of Pterospermum rubiginosum bark extract.

ETHNOPHARMACOLOGICAL RELEVANCE OF STUDY: Pterospermum rubiginosum is an evergreen plant in Western Ghats, India, used by traditional tribal healers due to its excellent biological potential for treating inflammation and pain relief procedures. The bark extract is also consumed to relieve the inflammatory changes at the bone fractured site. The traditional medicinal plant in India have to be characterized for its diverse phytochemical moieties, its interactive multiple target sites, and to reveal the hidden molecular mechanism behind the biological potency. AIM OF THE STUDY: The study focussed on plant material characterization, computational analysis (prediction study), toxicological screening (In vivo), and anti-inflammatory evaluation of P. rubiginosum methanolic bark extracts (PRME) in LPS-induced RAW 264.7 cells. MATERIALS AND METHODS: The pure compound isolation of PRME and their biological interactions were used to predict the bioactive components, molecular targets, and molecular pathways of PRME in inhibiting inflammatory mediators. The anti-inflammatory effects of PRME extract were evaluated in the lipopolysaccharide (LPS)-induced RAW264.7 macrophage cell model. The toxicity evaluation of PRME was performed in healthy 30 Sprague-Dawley experimental rats, were randomly divided into five groups for toxicological evaluation for 90 days. The tissue levels of oxidative stress and organ toxicity markers were measured using the ELISA method. Nuclear magnetic resonance spectroscopy (NMR) was performed to characterize the bioactive molecules. RESULTS: Structural characterization revealed the presence of vanillic acid, 4-O-methyl gallic acid, E-resveratrol, gallocatechin, 4'-O-methyl gallocatechin, and catechin. Molecular docking of NF-kB exhibited significant interactions with vanillic acid and 4-O-methyl gallic acid with binding energy -351.159 Kcal/Mol and -326.5505 Kcal/Mol, respectively. The PRME-treated animals showed an increase in total GPx and antioxidant levels (SOD and catalase). Histopathological examination revealed no variation in the liver, renal and splenic tissue's cellular pattern. PRME inhibited the pro-inflammatory parameters (IL-1ß, IL-6, and TNF-α) in LPS-induced RAW 264.7 cells. The protein level of TNF-α and NF-kB protein expression study brought out a notable reduction and exhibited a good correlation with the gene expression study. CONCLUSION: The current study establishes the therapeutic potential of PRME as an effective inhibitory agent against LPS-activated RAW 264.7 cells induced inflammatory mediators. Long-term toxicity evaluation on SD rats confirmed the non-toxic nature of PRME up to 250mg/body weight for 3 months.

Minerals, fatty acids, and antioxidant activity in sea knotgrass (Polygonum maritimum L.) seeds.

The design of the current study was to determine the functional properties of P. maritimum seeds by evaluating their mineral content, fatty acid composition, and biactive compounds. Results showed that seeds contained remarkable amounts of Na, K, Ca, Mg, Cu, Zn, and Fe. The oil yield was found to be 4.58% and contained higher unsaturated fatty acids predominated by linoleic and oleic acids. The colorimetric investigation revealed high total polyphenol (33.56 mg gallic acid equivalent/g), flavonoid (46.3 mg quercetin equivalent/g), and condensed tannin (22.3 mg catechin equivalent/g) contents. Furthermore, the LC-ESI/MS analysis revealed the presence of 13 phenolics mostly predominated by (+) catechin, quercetin-3-O-galactoside, gallic acid, and quinic acid. The seeds possessed strong antioxidant potential evidenced by ABTS and DPPH radical scavenging activities, total antioxidant capacity, and reducing power. The present findings suggest that P. maritimum seeds may be used as a potential source of biologically active natural products.

Effects of different antioxidants combined with high hydrostatic pressure on the color and anthocyanin retention of a blueberry juice blend during storage.

Blueberry juice has been found to undergo severe browning after treatment and cold storage, such as processing by high hydrostatic pressure (HHP) at 550 MPa/10 min/25 °C followed storage at 4 °C for 4 days. This browning may be due to the degradation of anthocyanin (AC) in the berries. Therefore, in this study, gallic acid (GA), ferulic acid (FA), ascorbic acid (VC), citric acid (CA), tea polyphenol (TP) and α-tocopherol (VE) were compared to determine their ability to improve the stability of the AC in HHP-treated blueberry juice. The juice was combined with the six abovementioned antioxidants at different concentrations, then treated by HHP at 550 MPa/10 min/25 °C and stored at 4 °C for 20 days. Thereafter, the pH levels, degrees °Brix, color parameters, total AC content and polyphenol oxidase (PPO) activity of the blueberry juice blend were measured and compared. Gallic acid at 2 g/L was found to be the most effective antioxidant to protect against AC degradation. After storage at 4 °C for 20 days, the AC content of the juice with no added antioxidants had decreased by 62.27% with a PPO relative activity of 50.78%, while the AC content of juice supplemented with 2 g/L GA had decreased by 13.42% with a PPO relative activity of 28.13%. The results of this study, thus, suggest that GA can stabilize the structure of AC in blueberry juice and reduce PPO activity, which may be beneficial in guiding the production of blueberry juice with high AC retention.

Polyphenolic Profile of Tunisian Thyme (Thymbra capitata L.) Post-Distilled Residues: Evaluation of Total Phenolic Content and Phenolic Compounds and Their Contribution to Antioxidant Activity.

During the last decade there has been growing interest in the formulation of new cosmetic, food and pharmaceutical products containing natural compounds with antioxidant activity and other beneficial properties. Aromatic and medicinal plants have always been the major source of bioactive compounds, especially, wild thyme (Thymbra capitata L.), which has been used since ancient times for its valuable health benefits that could be attributed to the richness of polyphenolic compounds. This study was undertaken with the following aims: to estimate the total polyphenolic content (TPC); to evaluate the antioxidant activity; to identify and quantify the phenolic compounds of post-distilled residues of Tunisian thyme, and their contribution to the antioxidant activity. The TPC, as determined by the Folin−Ciocalteu method, was found to reach the values of 126.7 and 107.84 mg gallic acid equivalent/g plant dry weight (mg GAE/g PDW). The antioxidant activity, which is assessed by DPPH and FRAP assays, reached the values of 42.97−45.64 µg/mL and 42.22−50.21 mMFe2+/mg PDW, respectively. HPLC analysis revealed the presence of fourteen polyphenolic compounds, of which diosmin and rosmarinic acid were found to be the most abundant (24.26 to 33.80 and 22.0.1 to 26.29 mg/g PDW, respectively). An important correlation was found between the antioxidant activity and several identified phenolic compounds (p < 0.05). The findings revealed that thyme post-distilled residues have an effective natural antioxidant potential due to their high concentration of bioactive molecules, and they appear to be useful in the pharmaceutical, cosmetic, and food industries, with beneficial effects on human health. Therefore, supplementing a balanced diet with herbs may have beneficial health effects.

The Ultrasound-Assisted Extraction of Polyphenols from Mexican Firecracker (Hamelia patens Jacq.): Evaluation of Bioactivities and Identification of Phytochemicals by HPLC-ESI-MS.

The objective of the present work was to optimize the extraction of phytochemicals from Hamelia patens Jacq. by ultrasound-assisted extraction. Taguchi L9 orthogonal array was used to evaluate the factors solid/liquid ratio (1:8, 1:12, and 1:16), extraction time (10, 20, and 30 min), and ethanol concentration (0, 35, and 70%). Total polyphenols were the response variable. Chromatographic fractionation using Amberlite XAD-16 was carried out and the total polyphenols, flavonoids, and condensed tannins were quantified. The redox potential, the reduction of the 2,2-diphenyl-1-picrylhydrazyl (DPPH), and the lipid oxidation inhibition were determined. Anti-bacterial activity was evaluated. The phytochemicals were identified by liquid chromatography coupled to mass spectrometry. Optimal extraction conditions were a solid/liquid ratio of 1:16, ethanol of 35%, and 10 min of ultrasound-assisted extraction. Maximum polyphenol content in the crude extract was 1689.976 ± 86.430 mg of gallic acid equivalents (GAE)/100 g of dried plant material. The purified fraction showed a total polyphenols content of 3552.84 ± 7.25 mg of GAE, flavonoids 1316.17 ± 0.27 mg of catechin equivalents, and condensed tannins 1694.87 ± 22.21 mg of procyanidin B1 equivalents, all per 100 g of purified fraction. Its redox potential was 553.93 ± 1.22 mV, reducing 63.08 ± 0.42% of DPPH radical and inhibiting 77.78 ± 2.78% of lipid oxidation. The polyphenols demonstrated antibacterial activity against Escherichia coli, Klebsiella pneumonia, and Enterococcus faecalis. The HPLC-ESI-MS analysis revealed the presence of coumarins, hydroxycinnamic acids, and flavonoids.

A Comparison of the Antiosteoporotic Effects of Cornelian Cherry (Cornus mas L.) Extracts from Red and Yellow Fruits Containing Different Constituents of Polyphenols and Iridoids in Osteoblasts and Osteoclasts.

Methods: Polyphenolic and iridoid constituents of extracts were analyzed qualitatively and quantitatively using the ultraperformance liquid chromatography system coupled with a quadrupole-time of flight mass spectrometry. Primary cultured osteoblasts isolated from mouse calvarias and osteoclast-lineage primary cultured monocytes isolated from mouse bone marrow were used for the assessment of osteoblast and osteoclast differentiation. In the osteoblast culture, cellular viability, alkaline phosphatase (ALP) activity, ALP staining, and mRNA expression of Alpl and Runx2 were examined. In the osteoclast culture, the examined parameters were cellular viability, tartrate-resistant acid phosphatase (TRAP) activity and staining, and mRNA expression of Nfatc1, Ctsk, and Acp. Results: A total of 41 main compounds of iridoids, anthocyanins, hydrolysable tannins, phenolic acids, and flavonols were identified in the three extracts. RED EXT1 contained most of the tested polyphenols and iridoids and was the only extract containing anthocyanins. YL EXT2 contained only one iridoid, loganic acid and gallic acid. YL EXT3 comprised a mixture of iridoids and polyphenols. RED EXT1, YL EXT 2, and to a lesser extent YL EXT3 promoted osteoblast differentiation increasing significantly ALP activity and the amount of ALP-positive stained cells. All extracts upregulated mRNA expression of Alpl and Runx2. RED EXT1 caused the most significant decrease in TRAP activity and the numbers of TRAP-positive multinucleated cells. RED EXT1 caused also the most significant downregulation of mRNA expression of osteoclast related genes Nfatc1, Ctsk, and Acp5. Extracts from yellow fruits, mostly YL EXT2 caused lower, but still significant inhibitory effect on TRAP and osteoclast related genes. Conclusions: The main conclusion of our study is that all three extracts, especially RED EXT1 from red cornelian cherry fruits, possess the antiosteoporotic potential and may be a promising phytomedicine candidate for the prevention and treatment of osteoporosis.

Potential of Kalanchoe pinnata as a Cancer Treatment Adjuvant and an Epigenetic Regulator.

Cancer is a global public health problem that is related to different environmental and lifestyle factors. Although the combination of screening, prevention, and treatment of cancer has resulted in increased patient survival, conventional treatments sometimes have therapeutic limitations such as resistance to drugs or severe side effects. Oriental culture includes herbal medicine as a complementary therapy in combination with chemotherapy or radiotherapy. This study aimed to identify the bioactive ingredients in Kalanchoe pinnata, a succulent herb with ethnomedical applications for several diseases, including cancer, and reveal its anticancer mechanisms through a molecular approach. The herb contains gallic acid, caffeic acid, coumaric acid, quercetin, quercitrin, isorhamnetin, kaempferol, bersaldegenin, bryophyllin a, bryophyllin c, bryophynol, bryophyllol and bryophollone, stigmasterol, campesterol, and other elements. Its phytochemicals participate in the regulation of proliferation, apoptosis, cell migration, angiogenesis, metastasis, oxidative stress, and autophagy. They have the potential to act as epigenetic drugs by reverting the acquired epigenetic changes associated with tumor resistance to therapy-such as the promoter methylation of suppressor genes, inhibition of DNMT1 and DNMT3b activity, and HDAC regulation-through methylation, thereby regulating the expression of genes involved in the PI3K/Akt/mTOR, Nrf2/Keap1, MEK/ERK, and Wnt/ß-catenin pathways. All of the data support the use of K. pinnata as an adjuvant in cancer treatment.

Antioxidant Capacity of Herzegovinian Wildflowers Evaluated by UV-VIS and Cyclic Voltammetry Analysis.

Considering the vast cultural and traditional heritage of the use of aromatic herbs and wildflowers for the treatment of light medical conditions in the Balkans, a comparison of the antioxidant capacity of wildflowers extracts from Herzegovina was studied using both cyclic voltammetry and spectrophotometry. The cyclic voltammograms taken in the potential range between 0 V and 800 mV and scan rate of 100 mV s-1 were used for the quantification of the electrochemical properties of polyphenols present in four aqueous plant extracts. Antioxidant capacity expressed as mmoL of gallic acid equivalents per gram of dried weight of the sample (mmoL GAE g-1 dw) was deduced from the area below the major anodic peaks (Q400 pH 6.0, Q500 pH 4.7, Q600 pH 3.6). The results of electrochemical measurements suggest that the major contributors of antioxidant properties of examined plants are polyphenolic compounds that contain ortho-dihydroxy-phenol or gallate groups. Using Ferric reducing-antioxidant power (FRAP) and 2,2'-azino-bis spectrophotometric methods (3-ethylbenzthiazoline-6-sulphonic acid) radical cation-scavenging activity (ABTS) additionally determined antioxidant capacity. The FRAP results ranged from 2.9702-9.9418 mmoL Fe/g dw, while the results for ABTS assays expressed as Trolox equivalents (TE) ranged from 14.1842-42.6217 mmoL TE/g dw. The Folin-Ciocalteu procedure was applied to determine the total phenolics content (TP). The TP content expressed as Gallic acid equivalents (GAE) ranged from 6.0343-9.472 mmoL GAE/g dw. The measurements of total flavonoid (TF) and total condensed tannin (TT) contents were also performed to obtain a broader polyphenolic profile of tested plant materials. Origanum vulgare L. scored the highest on each test, with the exception of TT content, followed by the Mentha × piperita L., Artemisia annua L., and Artemisia absinthium L., respectively. The highest TT content, expressed as mg of (-)catechin equivalents per gram of dried weight of sample (mg CE/g dw), was achieved with A. absinthium extract (119.230 mg CE/g dw) followed by O. vulgare (90.384 mg CE/g dw), A. annua (86.538 mg CE/g dw) and M. piperita (69.231 mg CE/g dw), respectively. In addition, a very good correlation between electrochemical and spectroscopic methods was achieved.

Chemical Composition and Bioctivity of the Giant Polypore or Black-Staining Mushroom, Meripilus giganteus (Agaricomycetes), from Serbia.

In this study, we tested the methanolic and acetonic extracts of the edible species Meripilus giganteus for their medicinal properties. Here, we present the chemical composition, as well as the antimicrobial, antioxidant, antidiabetic, neuroprotective, and cytotoxic activities, of the aforementioned extracts. The chemical composition of the extracts was analyzed with high-performance liquid chromatography coupled with diode-array detection and tandem mass spectrometry. The studied extracts contained 13 minerals, 23 polyphenols, 11 organic acids, and 22 carbohydrates. Antimicrobial activity was tested by applying the microdilution assay, and the results ranged from 0.002 to 20 mg/mL. Antioxidant activity was studied with the DPPH assay (half-maximal inhibitory concentration = 673.42 and 712.31 µg/mL for the methanolic and acetonic extracts, respectively) and the reducing power assay (range, 0.042-0.099 µg/mL). Total phenolic content was determined as 4.86 mg gallic acid equivalents (GAE)/g for the methanolic extract and 5.06 mg GAE/g for the acetonic extract. The antidiabetic effect of the extracts was explored with the α-amylase and α-glucosidase assays, and the results ranged from 30.66% to 38.67% and from 11.06% to 17.08%, respectively. The neuroprotective properties of M. giganteus extracts were studied using the acetylcholinesterase inhibition assay, with results ranging from 4.54% to 9.31%. Cytotoxic activity was tested by applying the microtetrazolium assay, with results ranging from 196.24 to 322.83 µg/mL. Although some assays proved relatively limited activities of the M. giganteus extracts studied, they can still be considered as natural sources of bioactive compounds without side effects.

Screening of potent α-glucosidase inhibitory and antioxidant polyphenols in Prunella vulgaris L. by bioreaction-HPLC-quadrupole-time-of-flight-MS/MS and in silico analysis.

Prunella vulgaris L. is a well-known traditional Chinese medicine for blood glucose homeostasis and antioxidant potential. Ethyl acetate fraction of P. vulgaris L. demonstrated higher phenolic content (85.53 ± 6.74 mg gallic acid equivalents per gram dry weight), α-glucosidase inhibitory (IC50 , 69.13 ± 2.86 µg/ml), and antioxidant (IC50 , 8.68 ± 1.01 µg/ml) activities. However, the bioactive polyphenols responsible for the beneficial properties remain unclear. Here, bioreaction-HPLC-quadrupole-time-of-flight-MS/MS method was developed for rapid, accurate, and efficient screening and identification of polyphenols with α-glucosidase inhibitory and antioxidant activities from P. vulgaris L. Bioactive polyphenols can specifically bind with α-glucosidase or react with 1,1-diphenyl-2-picryl-hydrazyl radical, which was easily discriminated from nonactive compounds. Subsequently, 20 bioactive polyphenols (16 phenyl propionic acid derivatives and four flavonoids) were screened and identified. Furthermore, molecular docking analysis revealed that screened 20 polyphenols bind with the active sites of α-glucosidase through hydrogen bonding and π-π stacking. Density functional theory calculations demonstrated their electron transport ability and chemical reactivity. The in silico analysis confirmed the screened results. In summary, this study provided a valuable strategy for rapid discovering bioactive compounds from complex natural products and offered scientific evidence for further development and application of P. vulgaris L.