Biological Properties of Recently Described Wild Bramble Rubus oklejewiczii against the Species from Similar Niches.

The aim of this study was to compare the biological properties, such as antiviral, antibacterial, and antioxidant, of recently described pentaploid species Rubus oklejewiczii with tetraploid taxa growing in similar habitats including R. plicatus, R. gracilis, and R. wimmerianus. The antiviral potential was analyzed against bacteriophages with different genetic material: phi6 (a surrogate for the SARS-CoV-2 virus), T7, phiX174, and MS2. Antibacterial properties of fruit and leaf extracts were determined against Staphylococcus aureus, Bacillus cereus, Escherichia coli, and Salmonella enterica. The total phenolic content, as well as anthocyanins, ascorbic acid, pH, and antioxidant properties (FRAP and DPPH) were determined. R. oklejewiczii leaf extract was characterized by the weakest antibacterial and antiviral properties, which was closely correlated with the lowest content of polyphenolic compounds and antioxidant properties. The strongest biological properties were observed for R. wimmerianus leaves. Fruit extracts were characterized by lower phenolic content and antioxidant activities than leaves, with the lowest values observed for R. oklejewiczii extract. The antibacterial properties of fruit extracts were strongest for R. gracilis. The strongest antiviral potential was observed for R. oklejewiczii and R. wimmerianus fruit extracts against the bacteriophage phi6, which correlated with the lowest pH and the highest ascorbic acid content. The positive effect of the higher ploidy of R. oklejewiczii for most of the analyzed biological properties was not observed except for the antiviral potential of fruit extract. Due to its large and tasty fruits, this species seems to be very promising for cultivation and attractive for consumers, even though most of its biological properties were not any better compared to other examined tetraploid species.

The Comparison of Honey Enriched with Laboratory Fermented Pollen vs. Natural Bee Bread in Terms of Nutritional and Antioxidant Properties, Protein In Vitro Bioaccessibility, and Its Genoprotective Effect in Yeast Cells.

The aim of the study was to compare the nutritional value and bioactivity of honey enriched with a 10% addition of natural bee bread and its substitutes obtained as a result of laboratory fermentation of bee pollen. Physicochemical parameters, antioxidant properties, as well as the bioaccessibility of proteins using an in vitro static digestion model were analyzed. The bioactivity of the obtained enriched honeys was tested using the yeast model. The research indicates the similarity of honeys with the addition of "artificial bee bread" to honey with natural ones. During in vitro digestion, good bioaccessibility of the protein from the tested products was demonstrated. The ability of the products to protect yeast cells against hydrogen superoxide-induced oxidative stress was demonstrated using a qualitative spot test, which was stronger in the case of enriched honey than in pure rapeseed control honey. Significant inhibition of the growth of both strains of yeast exposed to bee pollen-enriched honeys was also demonstrated. Furthermore, all tested samples showed significant genoprotective activity against the genotoxic effect of zeocin and the reduction of the number of DNA double-strand breaks by a minimum of 70% was observed.

Chemical Composition and Bioactivity of Laboratory-Fermented Bee Pollen in Comparison with Natural Bee Bread.

Bee bread is a valuable product obtained from the hive on a relatively small scale, while bee pollen is more easily available. Therefore, an effective laboratory method of converting pollen into a bee bread substitute is desired. The aim of the research was to verify the influence of selected factors (temperature, ultrasound) on the quality of obtained product using Lactobacillus rhamnosus inoculum. The composition of the fermented pollen was analyzed using Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES), Raman spectroscopy, and SDS-PAGE and compared to natural bee bread and the original pollen. In vitro biological activity was assessed as antioxidant activity using a yeast model (BY4741 and sod1∆ strains). Fermentation of pollen occurred spontaneously and after inoculation, as demonstrated by lower pH and higher lactic acid content. Raman spectroscopy and ICP-OES confirmed changes in composition compared to the initial pollen. Compared to bee bread, the fermented pollen showed a higher content of polyphenols and comparable antioxidant activity; moreover, it accelerated yeast growth rate. In addition, a protective effect was observed for Cu/Zn-superoxide dismutase 1 (sod1∆ yeast mutant exposed to hydrogen peroxide-induced oxidative stress). The higher fermentation temperature (25 °C) produces a more bee-bread-like product, while the use of ultrasound and starter culture seems to have no positive effect.

Assessing the Antimicrobial Properties of Honey Protein Components through In Silico Comparative Peptide Composition and Distribution Analysis.

The availability of reference proteomes for two honeybee species (Apis mellifera and Apis cerana cerana) opens the possibility of in silico studies of diverse properties of the selected protein fractions. The antimicrobial activity of honey is well established and related to its composition, including protein components. We have performed a comparative study on a selected fraction of the honey-related proteins, as well as other bee-secreted proteins, utilizing a publicly available database of established and verified peptides with antimicrobial properties. Using a high-performance sequence aligner (diamond), protein components with antimicrobial peptide sequences were identified and analyzed. The identified peptides were mapped on the available bee proteome sequences, as well as on model structures provided by the AlphaFold project. The results indicate a highly conserved localization of the identified sequences within a limited number of the protein components. Putative antimicrobial fragments also show high sequence-based similarity to the multiple peptides contained in the reference databases. For the 2 databases used, the lowest calculated percentage of similarity ranged from 30.1% to 32.9%, with a respective average of 88.5% and 79.3% for the Apis mellifera proteome. It was revealed that the antimicrobial peptides (AMPs) site is a single, well-defined domain with potentially conserved structural features. In the case of the examples studied in detail, the structural domain takes the form of the two ß-sheets, stabilized by α-helices in one case, and a six-ß-sheet-only domain localized in the C-terminal part of the sequence, respectively. Moreover, no significant differences were found in the composition of the antibacterial fraction of peptides that were identified in the proteomes of both species.

The Antioxidant, Antibacterial and Anti-Biofilm Properties of Rapeseed Creamed Honey Enriched with Selected Plant Superfoods.

The aim of the study is to evaluate the effect of the addition of selected fruits and herbs belonging to the "superfoods" category for the bioactivity of a rapeseed honey matrix. Flavored creamed honeys with nine types of various additives (2 and 4% of content) were prepared and analyzed for the content of total phenols, flavonoids, antioxidant (FRAP, DPPH and ABTS) and antibacterial activity against four strains of bacteria. Additionally, the impact of three months of storage on the antioxidant properties of the products obtained was examined. The significant dose-dependent increase in the content of bioactive ingredients and antioxidant capacity in spiced honeys, as compared to control honey, was observed. The highest enrichment was obtained for the addition of powdered sea buckthorn leaves and black raspberry fruits. Honey with the addition of sea buckthorn leaves inhibited the growth of P. aeruginosa, S. aureus and K. pneumonia, whereas honeys with black raspberry and blackcurrant fruits showed activity only on the latter two strains. Furthermore, what is more interesting, honey supplemented with sea buckthorn leaf and black raspberry fruits inhibited S. aureus biofilm formation at the sub-minimum inhibitory concentrations (sub-MICs), showing a dose-dependent anti-biofilm effect.

Tracking of Thermal, Physicochemical, and Biological Parameters of a Long-Term Stored Honey Artificially Adulterated with Sugar Syrups.

The growing phenomenon of honey adulteration prompts the search for simple methods to confirm the authenticity of honey. The aim of the study was to evaluate the changes in thermal characteristics, physicochemical parameters, antioxidant and enzymatic activity of honey subjected to artificial adulteration. Two series of products were prepared with the use of two different sugar syrups with an increasing dosage of adulterant (0 to 30%). After 24 months of storage, the quality of adulterated samples (partially crystallized) was assessed in comparison to the control honey (solid). Used adulteration changed physicochemical parameters and reduced antioxidant and enzymatic activity of honey (p < 0.05). The admixture of syrup and invert (p < 0.05) reduced the viscosity of liquid phase of delaminated honey in a dose-dependent manner. In the study, artificially adulterated honeys were controlled using the standard differential scanning calorimetry, DSC. In all adulterated honeys, a specific glass transition, TG, was observed in the range of 34-38.05 °C, which was not observed for control honey and pure adulterants. Moreover, the additional Tgs were observed in a wide range from -19.5 °C to 4.10 °C for honeys adulterated by syrup only. In turn, the Tg in range of 50.4-57.6 °C was observed only for the honeys adulterated by invert. These specific Tg seem to be useful to detect honey adulteration and to identify the kind of adulterant used.

SDS-PAGE Protein and HPTLC Polyphenols Profiling as a Promising Tool for Authentication of Goldenrod Honey.

The aim of the study was to use protein and polyphenolic profiles as fingerprints of goldenrod honey and to apply them for verification of the labeled variety. The markers for 10 honey samples were correlated with the standard physicochemical parameters and biological activity measured in vitro as antioxidant, antifungal and antibacterial activities. Honey proteins were examined regarding soluble protein, diastase and SDS-PAGE protein profile. The polyphenolic profile was obtained with the use of the HPTLC and the antioxidant activity was detected with standard colorimetric methods. The antimicrobial effect of representative honey samples of different chemical profiles was verified against E. coli and budding yeast. It was found that the SDS-PAGE technique allows for creating the protein fingerprint of the goldenrod honey variety which was consistent for 70% of tested samples. At the same time, the similarity of their polyphenolic profile was observed. Moreover, specific chemical composition resulted in higher bioactivity of honey against tested bacteria and yeast. The study confirmed the usefulness of both SDS-PAGE and HPTLC techniques in honey authentication, as an initial step for selection of samples which required pollen analysis.

Antiviral and Antibacterial Effect of Honey Enriched with Rubus spp. as a Functional Food with Enhanced Antioxidant Properties.

The aim of this study was to investigate the effect of blackberry and raspberry fruits (1 and 4%) and leaves (0.5 and 1%) on the biological activities of rape honey. Honey and plant material extracts were analyzed regarding total phenolic, flavonoid, anthocyanin contents, HPTLC and HPLC polyphenol profiles, as well as antioxidant activity. The antiviral potential was analyzed against bacteriophage phi 6-a coronavirus surrogate-whereas antimicrobial was tested against S. aureus and E. coli. Blackberry extracts were more abundant in antioxidants than raspberry extracts, with better properties found for leaves than fruits and for cultivated rather than commercial plants. The addition of both Rubus plant additives significantly increased the antioxidant potential of honey by four-fold (for 4% fruits additive) to five-fold (for 1% of leaves). Honey with the addition of fruits possessed higher antiviral potential compared with raw rape honey (the highest for 4% of raspberry fruit and 1% of blackberry leaf additive). Honey enriched with Rubus materials showed higher antibacterial potential against S. aureus than rape honey and effectively inhibited S. aureus biofilm formation. To summarize, honey enriched with Rubus fruit or leaves are characterized by increased pro-health value and can be recommended as a novel functional food.

The Use of HPTLC and SDS-PAGE Methods for Coniferous Honeydew Honey Fingerprinting Compiled with Mineral Content and Antioxidant Activity.

Fir honeydew honey is a uniquely beneficial product which is often subjected to adulteration; however, pollen analysis is not useful to verify this honey type. Fourteen samples of EU protected designation of origin fir honeydew honey gathered directly from apiaries were studied. Standards of legal requirements and additional parameters, i.e., specific optical rotation, mineral content, and antioxidant activity, were tested. Five nectar honeys of different varieties were used as a comparative material. HPTLC and SDS-PAGE methods were used to fingerprint the honey types. All honeys tested fulfilled the quality requirements in terms of water content, pH, total acidity, conductivity, HMF, and diastase number. They were defined as dark amber on the Pfund scale and exhibited positive specific rotation (+2.5 to 25). Honeydew honey surpassed the tested nectar honeys in terms of mineral content and antioxidant activity as well as total polyphenolic content, except for buckwheat honey. The sugar and polyphenolic profile obtained by HPTLC allowed to distinguish honeydew from nectar honeys. The same was achieved by SDS-PAGE protein profiling. Both techniques seem to be cheap and quick tools for precisely distinguishing honeydew honey.

The Study of Chemical Profile and Antioxidant Properties of Poplar-Type Polish Propolis Considering Local Flora Diversity in Relation to Antibacterial and Anticancer Activities in Human Breast Cancer Cells.

Nine samples of ethanolic extracts of poplar-type propolis (EEP) originated from South-Eastern Poland were analyzed in terms of the diversity of the flora around the apiary. The mineral composition, antioxidant properties, polyphenolic profile (HPTLC), and main polyphenolic constituents (HPLC-DAD) were determined. Only minor differences in chemical composition and antioxidant capacity between tested EEPs were found regardless of their botanical origin. However, the biological activity of the EEPs was more diversified. The tested EEPs showed stronger antibacterial activity against Gram-negative bacteria (Escherichia coli) compared to Gram-positive bacteria (Staphylococcus aureus and Staphylococcus epidermidis). Staphylococci biofilm inhibition occurred as a result of exposure to the action of four out of nine EEPs (P1-P4). Due to the various compositions of individual EEPs, a different MCF-7 cellular response was observed according to inhibition of cells migration and proliferation. Almost every sample inhibited the migration of breast cancer cells at a low concentration (0.04 µg/mL) of propolis. Even at the lowest concentration (0.02 µg/mL), each EEP inhibited the proliferation of MCF-7 cells, however, the level of inhibition varied between samples.

The enrichment of honey with Aronia melanocarpa fruits enhances its in vitro and in vivo antioxidant potential and intensifies its antibacterial and antiviral properties.

The effect of adding the chokeberry fruit additive to rape honey was studied with regard to the physicochemical properties and biological activity. Two samples of dried powdered fruits were used to enrich the honey (1 and 4% v/v) during creaming. The obtained products were characterized in terms of sugar content, acidity, conductivity, total phenolic, flavonoid and anthocyanin contents and HPTLC polyphenol profiles. The antioxidant properties of enriched honeys were studied in vitro (FRAP, DPPH, and ABTS) and in vivo using a S. cerevisiae model. The inhibitory effect against 5 bacterial strains and coronavirus surrogate bacteriophage phi6 was tested. The addition of chokeberry significantly modified the physicochemical properties of honey and enhanced its antioxidant potential (from 3 to 15 times). Using HPTLC analysis, the occurrence of flavonoids, phenolic acids, and anthocyanins in chokeberry enriched honey was determined. The modified honey protected yeast cells against H2O2-induced oxidative stress when used as a pretreatment agent. All tested bacteria were susceptible to enriched honey in a dose-dependent manner. The antiviral potential of enriched honey against the model bacteriophage was discovered for the first time. In terms of numerous health benefits determined, honey enriched with Aronia melanocarpa fruits can be considered as an interesting novel functional food, which may increase the consumption of chokeberry superfruits.

Searching for Differences in Chemical Composition and Biological Activity of Crude Drone Brood and Royal Jelly Useful for Their Authentication.

Drone brood is a little-known bee product which is frequently considered as a male equivalent of royal jelly and is sometimes used as its adulterant. The aim of the study was to compare the chemical composition and biological activity of both bee products originated from the same apiaries (n = 3) limiting the influence of genetic and environmental factors. Moreover, for drone brood study covered testing three stages of larval development (days 7, 11, and 14). The comparison included mineral composition (ICP-OES method), protein content and protein profile (SDS-PAGE), testosterone and estradiol content (ELISA tests). HPTLC method was used to analyze of sugar, amino acids, and polyphenolic profile of drone brood and royal jelly. Moreover, their antioxidant and enzymatic properties were compared. A lot of similarities between drone brood and royal jelly were found in terms of chemical components. However, drone brood was more abundant in iron and manganese, reducing sugars and some amino acids, especially proline, tyrosine, and leucine. It contained more testosterone (especially on the 14th day) and estradiol (on the 7th day). The greatest differences in the enzymatic activities and polyphenolic profile were found. Diastase and α-glucosidase activity were found as specific enzymes of the drone brood. Similarly, ferulic and ellagic acids were characteristic for brood and were not present in royal jelly. The study showed a lot of similar features for both tested bee products, however, some specific markers which can serve to differentiate drone brood and royal jelly were found.

The Comparison of Physicochemical Parameters, Antioxidant Activity and Proteins for the Raw Local Polish Honeys and Imported Honey Blends.

Many imported honeys distributed on the Polish market compete with local products mainly by lower price, which can correspond to lower quality and widespread adulteration. The aim of the study was to compare honey samples (11 imported honey blends and 5 local honeys) based on their antioxidant activity (measured by DPPH, FRAP, and total phenolic content), protein profile obtained by native PAGE, soluble protein content, diastase, and acid phosphatase activities identified by zymography. These indicators were correlated with standard quality parameters (water, HMF, pH, free acidity, and electrical conductivity). It was found that raw local Polish honeys show higher antioxidant and enzymatic activity, as well as being more abundant in soluble protein. With the use of principal component analysis (PCA) and stepwise linear discriminant analysis (LDA) protein content and diastase number were found to be significant (p < 0.05) among all tested parameters to differentiate imported honey from raw local honeys.

Antioxidant Activity of Frozen and Freeze-Dried Drone Brood Homogenate Regarding the Stage of Larval Development.

Drone brood is a little-known and poorly studied bee product used and valued in the treatment of many diseases, including male infertility and women's menopausal disorders. The aim of this study was to evaluate the antioxidant activity of drone brood depending on the stage of larval development and the method of preservation. Aqueous and ethanolic homogenate extracts of drone brood were assayed for antioxidant activity (with the DPPH, FRAP, and ABTS methods), polyphenol, and flavonoid content. The extracts' polyphenolic profiles were compared by the HPTLC method. Drone brood has been shown to be more active in the earlier stages of development (between days 7-11), with a decline in antioxidant activity in the later period (by the 14th day). The freeze-drying process did not cause significant changes in the antioxidant activity of brood preparations converted to dry mass. Based on the higher activity of the aqueous compared to 70% ethanolic extracts, it was shown that the dominant fraction of brood consisted of hydrophilic antioxidants. The results obtained with different methods were highly correlated, excluding those from the ABTS assay. The HPTLC method showed that the polyphenol fraction of drone brood homogenate consisted mainly of phenolic acids and flavonoids. It was shown that drone brood has valuable antioxidant properties that can be compared with royal jelly.

Caffeine content and antioxidant activity of various brews of specialty grade coffee.

BACKGROUND: The aim of the study was to evaluate the caffeine level and antioxidant activity of brews of specialty grade coffee compared to popular coffee brands. METHODS: Ten types of coffee were used, including 7 specialty Arabica, 1 Robusta and 2 popular cheap coffee brands. For caffeine determination, HPLC analysis and the spectrophotometric method were used as reference. The total polyphenol content and antioxidant capacity (DPPH and FRAP methods) were evaluated. For two selected high-quality coffees, the influence of the brewing method on the antioxidant activity and caffeine content in the brews was assayed. RESULTS: Regarding the caffeine content, differences between specialty coffee brews and popular products were not found, and an average level amounted to 56 and 40 mg/ml, respectively. In contrast, the antioxidant capacity of specialty coffee brews was significantly higher than for popular ones, independently of the test used. The highest scavenging ability and total phenolic content was found for S3 specialty coffee (46.15% of DPPH inhibition and 58.7 mg GAE/ml, respectively), whereas the lowest was found for popular coffee (about 35% of DPPH inhibition and about 41 mg/GAE/ml). For two selected high-quality coffees, the influence of the brewing method on the antioxidant activity and caffeine content in the brews was tested. It was found that the use of a dripper (overflow brewing method) provides the brew with the best antioxidant properties but with moderate caffeine levels. CONCLUSIONS: It was found that 'specialty' quality coffees do not differ from popular brands in terms of caffeine content, but significantly outweigh them in terms of antioxidant activity. This property can be beneficial in the case of a high consumption of coffee, due to antiradical protective effects without the risk of caffeine overdose.

Coffee Extends Yeast Chronological Lifespan through Antioxidant Properties.

Aging is a multifactorial process accompanied by loss of cell function. Science has been looking for factors responsible for aging for many years. However, despite identifying a number of possible causes, the definite reason for aging has been elusive so far. One of the factors contributing to aging is oxygen free radicals. In this context, beneficial effects of coffee on various organisms, including humans, were investigated, although the results are far from unequivocal. In our research, we used the budding yeast-something of a workhorse in many studies, including the studies of aging. So far, the impact of coffee on the aging of cells in the budding yeast experimental setup has little known about it. Here, we provide strong evidence that coffee compounds, particularly flavonoids, are responsible for scavenging free radicals and longevity in yeast lacking Sod1, Sod2 and Rad52 proteins. In this paper, we compared Arabica and Robusta coffee types. We present an analysis of the concentration of caffeine and flavonoids measured by the High-Performance Liquid Chromatography method. We show that Robusta has a much greater antioxidant capacity than Arabica. We also conclude that coffee infusions significantly extend the chronological lifespan of the Saccharomyces cerevisiae yeast cells by protecting cells against reactive oxygen species, double DNA-strand break and decrease in metabolic activity.

Analysis of Cytotoxicity of Selected Asteraceae Plant Extracts in Real Time, Their Antioxidant Properties and Polyphenolic Profile.

Plants from Asteraceae family are widely used for their therapeutic effects in the treatment of gastrointestinal diseases, but the consequences of excessive intake still need to be studied. The aims of this study were the evaluation of cytotoxicity, measurement of antioxidant properties and determination of polyphenolic profile of Tanacetum vulgare L. (tansy), Achillea millefolium L. (yarrow) and Solidago gigantea Ait. (goldenrod) ethanolic extracts. The cytotoxicity of extracts was monitored by xCELLigence system in real time by using porcine intestinal epithelial cell line (IPEC-1) and by measurement of changes in metabolic activity ((3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) (MTS) assay). The antioxidant properties were measured by spectrophotometric methods and polyphenolic profiles were determined by HPLC-DAD for 50% ethanol extracts (10% w/v). Strong cytotoxic effect was recorded for tansy and yarrow extracts (125-1000 µg/mL) by xCELLigence system and MTS assay. Conversely, a supportive effect on cell proliferation was recorded for goldenrod extracts (125 µg/mL) by the same methods (p < 0.001). The antioxidant activity was in good correlation with total polyphenolic content, and the highest value was recorded for goldenrod leaves, followed by tansy leaves, goldenrod flowers and yarrow leaf extracts. The goldenrod extracts were abundant with flavonoids, whereas phenolic acid derivatives predominated in the polyphenolic profile of tansy and yarrow.

The Effect of Adding the Leaves and Fruits of Morus alba to Rape Honey on Its Antioxidant Properties, Polyphenolic Profile, and Amylase Activity.

Two various species of mulberry (Morus sp.) were selected to enrich rape honey with dried leaves or lyophilized fruits (4% w/v). Finally, fruits and leaves of the 'Ukrainska' clone were introduced into the honey during creaming in concentrations from 1 to 4% w/v. The total phenolic content, antioxidant activity, anthocyanins content, and polyphenolic profile were tested in plant extracts and enriched honeys. Moreover, α-glucosidase, ß-galactosidase, and diastase activities were investigated in honeys. For mulberry extracts, chlorogenic acid isomers and rutin were considered main antioxidant compounds. The antioxidant activity of honey enriched with mulberry leaves increased even more than 50 times, due to introducing numerous phenolic acids and flavonoid glycosides. A significant decrease in the diastase activity in honey depending on the content of added mulberry leaves (almost 50% decrease in the case of 4% addition) was found, suggesting the inhibitory effect of honey with mulberry leaves against carbohydrate hydrolyzing enzymes.

Polyphenols Content, Antioxidant Activity, and Cytotoxicity Assessment of Taraxacum officinale Extracts Prepared through the Micelle-Mediated Extraction Method.

This experiment was conducted with extracts prepared from dandelion (Taraxacum officinale F. H. Wigg) leaves and flowers, using the micelle-mediated extraction method, with the surface active compound Triton X-100 and water⁻acetone as the extraction solvents. Extracts were, first, examined for the content of total phenols and the antioxidant capacity. All extracts showed good anti-radical properties, especially for leaves, in comparison to the flower samples. Flavonoids (mainly luteolin derivatives) and phenolic acids, predominated among the determined polyphenols. Quantitative analyses indicated acetone extract to be the richest in phenols (up to 0.535 mg/mL), in the case of dandelion leaves, and Triton X-100 extract in the case of flowers (0.385 mg/mL). Extracts were also evaluated for cytotoxicity to the model cell line (epithelial rabbit kidney cells RK13), using the colorimetric 3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide (MTT) test and the real-time cell analysis method ((RTCA); xCELLigence system). The obtained results indicated that surfactants, especially non-ionic ones, can be effectively used as modifiers in the aqueous extraction of phenolic compounds from plant materials. An advantage over the traditional organic solvents is their non-flammability. Furthermore, surfactants might also be used at low concentrations. Studies on cell lines, however, indicated the cytotoxic effect of this type of compound, even in the trace amounts present in the extracts.

The Chemical Composition of Achillea wilhelmsii C. Koch and Its Desirable Effects on Hyperglycemia, Inflammatory Mediators and Hypercholesterolemia as Risk Factors for Cardiometabolic Disease.

This study was done to identify the content compounds of Achillea wilhelmsii (A. wilhelmsii) and to evaluate its hypoglycemic and anti-hypercholesterolemic activity and effect on inflammatory mediators. The extracts and fractions of A. wilhelmsii were thoroughly analyzed using high performance liquid chromatography (HPLC), and the total content of phenols and flavonoids was determined. The hypoglycemic activity was evaluated in vivo using alloxan-induced diabetic mice. The effect upon inflammatory mediators was evaluated in vitro using the human monocytic leukemia cell line (THP-1). The anti-hypercholesterolemic activity was evaluated in vitro using the 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase assay kit. The water extract (WE)-treated group showed the highest reduction in the fasting blood glucose levels (FBGL). The chloroform fraction (CF) and ethyl acetate fraction (EAF) both showed a significant ability to reduce the secretion of tumor necrosis factor alpha (TNF-α). The EAF, however, also attenuated the levels of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9). The CF showed the most significant 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) inhibition activity. The five main compounds in the CF were isolated and identified. Out of the five compounds in the CF, 1ß,10ß-epoxydesacetoxymatricarin (CP1) and leucodin (CP2) showed the highest anti-hypercholesterolemic potential. A molecular docking study provided corresponding results.