Phytochemical screening of Pulsatilla species and investigation of their biological activities

© 2019, The Author(s). We previously demonstrated that extracts from Echinacea purpurea material varied substantially in their ability to activate macrophages in vitro and that this variation was due to differences in their content of bacterial components. The purpose of the current study was to identify soil conditions (organic matter, nitrogen, and moisture content) that alter the macrophage activation potential of E. purpurea and determine whether these changes in activity correspond to shifts in the plant-associated microbiome. Increased levels of soil organic matter significantly enhanced macrophage activation exhibited by the root extracts of E. purpurea (p \u3c 0.0001). A change in soil organic matter content from 5.6% to 67.4% led to a 4.2-fold increase in the macrophage activation potential of extracts from E. purpurea. Bacterial communities also differed significantly between root materials cultivated in soils with different levels of organic matter (p \u3c 0.001). These results indicate that the level of soil organic matter is an agricultural factor that can alter the bacterial microbiome, and thereby the activity, of E. purpurea roots. Since ingestion of bacterial preparation (e.g., probiotics) is reported to impact human health, it is likely that the medicinal value of Echinacea is influenced by cultivation conditions that alter its associated bacterial community

Chemical composition, antimicrobial, and antioxidant activities of the essential oils from stem, leaves, and seeds of Caryopteris foetida (D. don) Thell.

214-224Caryopteris foetida (D. Don) Thell. also known as stinking blue beard is an aromatic shrub of family verbenaceae. The essential oils from the leaf, stem, and seeds of C. foetida were extracted and analyzed in search of a novel compound. Over 73 constituents were identified from the essential oils of leaf, seeds, and stem. The major compound identified in the leaf oil was δ-cadinene (15.4%) followed by β-caryophyllene (7.8%), (E)-β-farnesene (8.3%), γ-cadinene (7.5%), spathulenol (7.2%), and τ-muurolol (5.1%). The stem oil predominantly consisted of methyl 7-methylcyclopenta[c]pyran-4-carboxylate (15.8%) along with a significant amount of δ-cadinene (11.6%) and γ-cadinene (5.6%), whereas seed oil was characterized by the presence of β-caryophyllene (14.3%) along with α-humulene (8.6%) and β-bisabolene (9.4%). The essential oils were screened for their in vitro antioxidant potential in terms of radical scavenging, metal chelating and reducing assay. The leaf oil exhibited strong DPPH radical scavenging (IC50 =5.1±0.2 μg/mL) and reducing activity (IC50 =3.6±0.5 μg/mL). The antibacterial potential was tested against Salmonela typhimurium, Escherichia coli, Staphylococcus Aureus, and Bacillus megaterium. Maximum activity was exhibited by the essential oils from the leaf and seed. The essential oils were also analyzed for their in vitro anti-inflammatory activity by the protein denaturation method. Both the leaf and stem essential oils exhibited good anti-inflammatory activity with IC50 value of 12.8±0.0 μg/mL and 17.3±0.0 μg/mL respectively

"Heterobasidion annosum" induces apoptosis in DLD-1 cells and decreases colon cancer growth in In vivo model

Application of substances from medicinal mushrooms is one of the interesting approaches to improve cancer therapy. In this study, we commenced a new attempt in the field of Heterobasidion annosum (Fr.) Bref. sensu lato to further extend our knowledge on this basidiomycete fungus. For this purpose, analysis of the active substances of Heterobasidion annosum methanolic extract and also its influence on colorectal cancer in terms of in vitro and in vivo experiments were performed. In vivo studies on mice were conducted to verify its acute toxicity and to further affirm its anticancer potential. Results indicated that all the most common substances of best known medicinal mushrooms that are also responsible for their biological activity are present in tested extracts. In vitro tests showed a high hemocompatibility and a significant decrease in viability and proliferation of DLD-1 cells in a concentration-dependent manner of Heterobasidion annosum extract. The studies performed on xenograft model of mice showed lower tendency of tumor growth in the group of mice receiving Heterobasidion annosum extract as well as mild or moderate toxicity. Obtained results suggest beneficial potential of Heterobasidion annosum against colon cancer as cytotoxic agent or as adjuvant anticancer therapy

Phytochemical screening of Pulsatilla species and investigation of their biological activities

The present study aimed to identify biologically active secondary metabolites from the rare plant species, Pulsatilla patens subsp. patens and the cultivated P. vulgaris subsp. vulgaris. Chromatographic fractionation of the ethanolic extract of the roots of P. patens subsp. patens resulted in the isolation of two oleanane-type glycosides identified as hederagenin 3-O-β-d-glucopyranoside (2.7 mg) and hederagenin 3-O-β-d-galactopyranosyl-(1→2)-β-d-glucopyranoside (3.3 mg, patensin). HPLC analysis of the methanolic extract of the crude root of P. patens subsp. patens and P. vulgaris subsp. vulgaris revealed the presence of Pulsatilla saponin D (hederagenin 3-O-α-l-rhamnopyranosyl(1→2)-[β-d-glucopyranosyl(1→4)]-α-l-arabinopyranoside). Chromatographic analysis using GC-MS of the silylated methanolic extracts from the leaves and roots of these species identified the presence of carboxylic acids, such as benzoic, caffeic, malic, and succinic acids. The extracts from Pulsatilla species were tested for their antifungal, antimicrobial, and antimalarial activities, and cytotoxicity to mammalian cell lines. Both P. patens subsp. patens and P. vulgaris subsp. vulgaris were active against the fungus Candida glabrata with the half-maximal inhibitory concentration (IC50) values of 9.37 µg/mL and 11 µg/mL, respectively. The IC50 values for cytotoxicity evaluation were in the range of 32–38 μg/mL for P. patens subsp. patens and 35–57 μg/mL for P. vulgaris subsp. vulgaris for each cell line, indicating general cytotoxic activity throughout the panel of evaluated cancer and noncancer cells

Research on the chemical composition of birch (Betula L.) buds

W ramach pracy doktorskiej przeprowadzono szczegółowe badania składu chemicznego pąków brzozy brodawkowatej (B. pendula) oraz brzozy omszonej (B. pubescens), które są najbardziej rozpowszechnionymi na kontynencie eurazjatyckim gatunkami brzozy. Wyciągi z pąków B. pendula oraz B. pubescens pozyskano stosując pięcioetapową ekstrakcję sekwencyjną, w której użyto kolejno: CO2 w stanie nadkrytycznym (scCO2), n-heksanu, chloroformu, eteru dietylowego oraz mieszaniny metanol:woda (4:1). Przeprowadzono szczegółową analizę składu chemicznego uzyskanych wyciągów za pomocą GC-MS. W wyniku przeprowadzonych rozdziałów chromatograficznych wydzielono trzy związki triterpenowe, których struktury określono przy użyciu spektroskopii 1H oraz 13C NMR. Wyizolowane związki, tj. kwas 3,4-seko-damara-4(29),20(21),24(25)-trien-3-owy, kwas 20-hydroksy-3,4-seko-damara-4(29),24(25)-dien-3-owy i kabraleon, zostały przekazane do badań ich aktywności antynowotworowej. Dokonano również ilościowego oznaczenia wybranych flawonoidów oraz triterpenów w wyciągach scCO2 z pąków brzóz. Ponadto przeprowadzono analizy substancji lotnych emitowanych przez pąki 22 gatunków brzozy. Na podstawie otrzymanych wyników badań wyróżniono trzy grupy brzóz. Wykazano, że uzyskane wyniki badań mogą być wykorzystane do celów chemosystematyki rodzaju Betula.In the doctoral dissertation, a detailed chemical composition study of buds from silver birch (B. pendula) and downy birch (B. pubescens) was decided to be conducted, which are the most widespread species of a birch on the Eurasian continent. Extracts from B. pendula and B. pubescens buds were obtained using five-step sequential extraction, using successively: supercritical CO2 (scCO2), n-hexane, chloroform, diethyl ether, and methanol: water (4: 1). The analysis of chemical compositions of extracts was performed using GC-MS. As a result of the chromatographic separations, three triterpenes were isolated, and the structures of these compounds were determined using 1H and 13C NMR spectroscopy. Isolated compounds, i.e. 3,4-seco-dammara-4(29),20(21),24(25)-trien-3-oic acid, 20-hydroxy-3,4-seco-dammara-4(29),24(25)-dien-3-oic acid and cabraleone were further subjected to research of their anti-cancer activity. Furthermore, the quantitative determinations of flavonoids and selected triterpenes in scCO2 extracts from birch buds were performed. Moreover, the analysis of volatiles emitted from buds of 22 species of birch were realized. Based on obtained research results, three groups of birches were distinguished. It was revealed, that the results of this research can be useful for chemosystematic purposes of Betula genus.Uniwersytet w Białymstoku. Wydział Chemii

GC-MS investigation of the chemical composition of honeybee drone and queen larvae homogenate

Honeybee larva homogenate appears to be underrated and insufficiently explored but this homogenate is an exceptionally valuable honeybee product. Drone larva homogenate is very nutritional due to its high content of proteins, free amino acids, lipids, and carbohydrates. Moreover, the biological characteristics of honeybee larvae indicate the presence of chemical substances that may be pharmacologically active. In spite of the above, the chemical composition of honeybee larva has not gained as much attention as that of other bee products. In this study, the chemical composition of honeybee brood homogenate has been investigated using gas chromatography/mass spectrometry. As a result, it was possible to isolate as many as 115 extractive organic compounds from 6 samples of crude queen and 9 samples of drone homogenate. The main groups of substances extracted from either type of homogenate were composed of free amino acids and carbohydrates. The relative content of amino acids in queen homogenate as well as the share of essential amino acids were found to be higher than in the drone homogenate. Disaccharide trehalose was the dominant sugar in the queen larvae, whilst glucose prevailed in the drone larvae. Comparative chemical analyses of honeybee queen and drone larva homogenates have allowed us to make a preliminary inference about a higher overall value of the former

Cytotoxicity of Triterpene Seco-Acids from Betula pubescens Buds

The present study investigated the magnitude and mechanism of the cytotoxic effect on selected cancer cell lines of 3,4-seco-urs-4(23),20(30)-dien-3-oic acid (1), 3,4-seco-olean-4(24)-en-19-oxo-3-oic acid (2), and 3,4-seco-urs-4(23),20(30)-dien-19-ol-3-oic acid (3) isolated from downy birch (Betula pubescens) buds by carbon dioxide supercritical fluid extraction and gradient column chromatography. Cell viability in six human cancer lines exposed to these compounds was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Apoptosis was quantified by annexin V/propidium iodide staining of gastric cancer AGS and colorectal cancer DLD-1 cells. To evaluate the mechanism of apoptosis, the expression of apoptosis-related proteins was analyzed by Western blot. Compound 1 exhibited non-specific toxicity, while compounds 2 and 3 were specifically toxic to colon and stomach cancer cells. The toxicity of compounds 2 and 3 against these two cell lines was greater than for compound 1. Cleavage of caspase-8, -9, and -3 was found in AGS and DLD-1 cells treated with all three seco-acids, indicating the induction of apoptosis via extrinsic and intrinsic pathways. Therefore, triterpene seco-acids (1–3) decreased cell viability by apoptosis induction. AGS and DLD-1 cells were more susceptible to seco-acids with an oxidized C19 than normal fibroblasts. Hence, it made them a new group of triterpenes with potential anticancer activity

Parametric Modelling of the Crystalline Microstructure of the MCM41-Type Mesoporous Silica Modified with Derivatives of Alkyls

A siliceous material in which a framework order was established with a surfactant with sixteen carbon atoms in alkyl chains, MCM-41-C16, was synthesised, surface-modified, and tested regarding the selected physical properties. The pristine material was extracted in an acidic aqueous alcohol and then lined with different surface groups. The properties of four adsorbents were investigated using XRD, X-ray photoelectron spectroscopy, and N2 physisorption techniques. The unit–cell constant was determined from X-ray diffractograms, being in fixed relation to the edge length of the hexagonal frame. The specific surface areas of mesopores and whole crystallites were determined from low-temperature N2-physisorption isotherms. The novelty of this work is a mathematical model of a crystalline microstructure explaining the sizes and shapes of crystalline grains in relation to adsorption features, proposed and successfully tested with the aforementioned experimental data. The roughness of the surface is different from one that is necessary to explain the experimental characteristics quantitatively

Chemical composition, antimicrobial, and antioxidant activities of the essential oils from stem, leaves, and seeds of Caryopteris foetida (D. don) Thell.

Caryopteris foetida (D. Don) Thell. also known as stinking blue beard is an aromatic shrub of family verbenaceae. The essential oils from the leaf, stem, and seeds of C. foetida were extracted and analyzed in search of a novel compound. Over 73 constituents were identified from the essential oils of leaf, seeds, and stem. The major compound identified in the leaf oil was δ-cadinene (15.4%) followed by β-caryophyllene (7.8%), (E)-β-farnesene (8.3%), γ-cadinene (7.5%), spathulenol (7.2%), and τ-muurolol (5.1%). The stem oil predominantly consisted of methyl 7-methylcyclopenta[c]pyran-4-carboxylate (15.8%) along with a significant amount of δ-cadinene (11.6%) and γ-cadinene (5.6%), whereas seed oil was characterized by the presence of β-caryophyllene (14.3%) along with α-humulene (8.6%) and β-bisabolene (9.4%). The essential oils were screened for their in vitro antioxidant potential in terms of radical scavenging, metal chelating and reducing assay. The leaf oil exhibited strong DPPH radical scavenging (IC50 =5.1±0.2 µg/mL) and reducing activity (IC50 =3.6±0.5 µg/mL). The antibacterial potential was tested against Salmonela typhimurium, Escherichia coli, Staphylococcus Aureus, and Bacillus megaterium. Maximum activity was exhibited by the essential oils from the leaf and seed. The essential oils were also analyzed for their in vitro anti-inflammatory activity by the protein denaturation method. Both the leaf and stem essential oils exhibited good anti-inflammatory activity with IC50 value of 12.8±0.0 µg/mL and 17.3±0.0 µg/mL respectively

The Differentiation of the Infestation of Wheat Grain with <i>Fusarium poae</i> from Three Other <i>Fusarium</i> Species by GC–MS and Electronic Nose Measurements

The massive import of uncontrolled technical grain from the East into the European Community poses a risk to public health when it ends up in the mills to be used as flour for food purposes instead of being burnt (biofuel). In fungal infections of wheat, the most dangerous species belong to the genus Fusarium. F. poae is a pathogen that is most commonly isolated from cereals worldwide and causes various types of diseases in animals and humans due to the numerous toxins it produces. The manuscript reports an attempt to distinguish between four species of Fusarium, F. avanceum, F. langsethiae, F. poae, and F. sporotrichioides, in wheat grains by measuring the volatiles emitted. The patterns obtained from the signals captured by the electronic nose PEN3 were used to build the Random Forests classification model. The recall and precision of the classification performance for F. poae reached 91 and 87%, respectively. The overall classification accuracy reached 70%. Gas chromatography coupled with mass spectrometry (GC–MS) was used to analyze the chemical composition of the emitted volatiles. The patterns found in the GC–MS results allowed an explanation of the main patterns observed when analyzing the electronic nose data. The mycotoxins produced by the Fusarium species analyzed were detected. The results of the reported experiment confirm the potential of the electronic nose as a technology that can be useful for screening the condition of the grain and distinguishing between different pathogenic infestations