Ailanthus altissima Seed Oil-A Valuable Source of Lipid-Soluble Components with DNA Protective and Antiproliferative Activities.

The present study is focused on the chemical and lipid composition of seed oil of the European ornamental and invasive wood plant Ailanthus altissima (Simaroubaceae). Total lipids, proteins, carbohydrates, ash, and moisture in the seeds were determined. A high yield of glyceride oil (30.7%) was found, as well as a high content of fibers (29.6%) and proteins (18.7%). Physicochemical properties of the oil define it as semi-dry (129.4 g I2/100 g Iodine value) with oxidative stability, refractive index, saponification value, and relative density similar to widely used oils with nutritional value and health benefits. The composition of the seed oil was determined chromatographically. Unsaturated fatty acids (95.3%) predominated in the seed oil, of which linoleic acid (48.6%) and oleic acid (44.8%) were the major ones. The main lipid-soluble bioactive components were ß-sitosterol (72.6%), γ-tocopherol (74.6%), phosphatidylinositol (29.5%), and phosphatidic acids (25.7%). The proven in vitro DNA-protective ability of seed oil is reported for the first time. The seed oil exhibited a weak antiproliferative effect on HT-29 and PC3 tumor cell lines and showed no cytotoxicity on the BALB/c 3T3 cell line. In brief, the present study reveals that A. altissima seed oil can be used as a healthy food.

Chemical Composition, Lipid-Soluble Bioactive Compounds and Potential Health Benefits of the Moss Hypnum cupressiforme Hedw.

Hypnum cupressiforme Hedw. is the main species for Moss surveys (ICP Vegetation programme) in Southeastern Europe and is widely distributed in the region. In addition to their biomonitoring role, mosses are applied in some countries as a traditional medicine for the treatment of eczema, cuts, burns, eye diseases, etc. Therefore, the chemical and lipid composition of the moss H. cupressiforme is of interest to establish their possible application in different fields. The chemical composition of the moss was examined regarding total lipids, proteins, carbohydrates (i.e., fibres), ash, and moisture content. The main lipid-soluble bioactive components were determined as sterols, tocopherols, phospholipids and fatty acids. The major fatty acids were linoleic (14.9%), oleic (13.8%), palmitic (12.5%) and α-linolenic (11.3%) acids. Unsaturated fatty acids (56.4%) prevailed in the glyceride oil, in which the polyunsaturated ones constituted 32.5%. The lipid indices (atherogenicity, thrombogenicity, hypocholesterolemic/hypercholesterolemic ratio, peroxidability, and oxidation stability index) were also theoretically calculated based on the fatty acid composition of the moss lipids to establish their health benefits and the rate of oxidation. The primary results of this study revealed H. cupressiforme to be a promising alternative source of bioactive compounds that could be implemented in supplements with health-promoting effects.

Phenolic Profile, Antioxidant and DNA-Protective Capacity, and Microscopic Characters of Ailanthus altissima Aerial Substances.

Invasive species as sources of natural components are of increasing interest for scientific research. This is the case of Ailanthus altissima, which belongs to the top 100 of the most dangerous invasive plant species in Europe, and which is the subject of the present study. The purpose of the research was to analyze the main phenolic compounds in the flowers, leaves, and stem bark of A. altissima and determine the DNA-protective and antioxidant potential of their ethanolic extracts. HPLC profiling revealed the presence of 6 flavonoids and 10 phenolic acids, of which 15 were found in flowers, 14 in leaves, and 11 in the stem bark. Rutin (5.68 mg/g dw in flowers), hesperidin (2.67 mg/g dw in leaves) and (+)-catechin (2.15 mg/g dw in stem bark) were the best-represented flavonoids. Rosmarinic (10.32 mg/g dw in leaves) and salicylic (6.19 mg/g dw in leaves) acids were predominant among phenolic acids. All plant extracts tested showed in vitro antioxidant activity (determined by DPPH, ABTS, FRAP, and CUPRAC assays) and DNA-protection capacity (assay with supercoiled plasmid DNA-pUC19). The highest antioxidant activity was recorded in the flower parts (in the range from 661 to 893 mmol TE/g dw), followed by the leaves. A DNA protective potential for A. altissima leaf and flower extracts has not been established to date. In addition, the main microscopic diagnostic features of studied plant substances were described, with data for the flower parts being reported for the first time. The present study proves that A. altissima could be a natural source of DNA protection and antioxidants.

Koelreuteria paniculata Seed Oil-A Rich Natural Source of Unsaturated Fatty Acids and Phytocompounds with DNA Protective Potential.

The present work is focused on the physicochemical characteristics, chemical composition, and some biological activities of Koelreuteria paniculata seed oil. The glyceride oil, obtained with a Soxhlet apparatus by extraction with hexane, was characterized by a relatively high oil content (over 20%), and it is defined as a non-drying oil (iodine value-44 gI2/100 g) with good oxidative stability (over 50 h). There were identified 11 fatty acids, 6 sterols, 3 tocopherols, and 6 phospholipids, as the last group was reported for the first time. The major components among them were-monounsaturated eicosenoic and oleic acids, ß-sitosterol, ß-tocopherol, and phosphatidylcholine. The in vitro tests demonstrated DNA protective activity and a lack of cytotoxicity of the oil, data that has been reported for the first time. The in vitro MTT test of the oil on HT-29 and PC3 cell lines did not indicate antitumor activity. The seed oil studied contains valuable bio-components, which have proven benefits for human health, and that is why it could be used in food, cosmetic, and pharmaceutical products.

Antioxidant and DNA-Protective Potentials, Main Phenolic Compounds, and Microscopic Features of Koelreuteria paniculata Aerial Parts.

Interest in plant extracts as a natural source of antioxidants has grown significantly in recent years. The tree species Koelreuteria paniculata deserves attention due to its wide distribution, good adaptability, and growth to the degree of invasiveness in a number of European countries. The purpose of the present study was to analyze flavonoids and phenolic acids of the ethanol extracts from aerial parts of K. paniculata and to screen their antioxidant and DNA-protective activity. HPLC profiling revealed the presence of five flavonoids, with rutin (4.23 mg/g DW), hesperidin (2.97 mg/g DW), and quercetin (2.66 mg/g DW) as the major ones in the leaves, and (-)-epicatechin (2.69 mg/g DW) in the flower buds. Among the nine phenolic acids identified, rosmarinic, p-coumaric, salicylic, vanillic, and gallic acids were the best represented. All the extracts tested showed in vitro antioxidant activity that was determined by DPPH, ABTS, FRAP, and CUPRAC assays. The highest activity was recorded in the flower parts (in the range from 1133 to 4308 mmol TE/g DW). The DNA-protective capacity of the flower and stem bark extracts from the in vitro nicking assay performed, as well as the main diagnostic microscopic features of the plant substances, are given for the first time. According to the results obtained, the aerial parts of K. paniculata could be valuable sources of natural antioxidants.

Chemical Composition and Antimicrobial Activity of Essential Oil of Fruits from Vitex agnus-castus L., Growing in Two Regions in Bulgaria.

The chemical composition of Vitex agnus-castus L. (Verbenaceae family) fruits, collected from two regions in Bulgaria (south-central and north-east Bulgaria), was investigated. The content of proteins (5.3-7.4%), carbohydrates (73.9-78.8%), fiber (47.2-49.9%), ash (2.5-3.0%), essential oils (0.5%), and vegetable oil (3.8-5.0%) were identified in the fruits. The composition of the essential oils (EOs) of Vitex fruits from both regions was determined; the main compounds were 1,8-cineole (16.9-18.8%), α-pinene (7.2-16.6%), sabinene (6.7-14.5%), and bicyclogermacrene (7.3-9.0%), but significant differences in the quantitative and qualitative composition of EOs between the regions were found. The EOs of plants from north-east Bulgaria demonstrated antimicrobial activity against the pathogenic species Salmonella abony, Staphylococcus aureus, and Bacillus subtilis, but the Gram-negative bacteria Esсherichia coli and Pseudomonas aeruginosa exhibited resistance to the oil. Linoleic acid predominated in vegetable oil from both regions, followed by oleic acid. ß-sitosterol and γ-tocopherol were the main components in the sterol and tocopherol fraction of the lipids. Phosphatidic acids were the main components in the vegetable oil from north-east Bulgaria, while in the vegetable oil from south-central Bulgaria, all phospholipids were found in almost the same quantity. Overall, significant differences were observed in the chemical composition (proteins, carbohydrates, ash and moisture) of the fruits from the two regions of Bulgaria, as well as in the content of the main components of their essential and vegetable oils.

Chemical Compounds, Antitumor and Antimicrobial Activities of Dry Ethanol Extracts from Koelreuteria paniculata Laxm.

Koelreuteria paniculata Laxm. is used in traditional medicine and has various established biological activities, however, the species is considered to be a potentially invasive alien tree species for Bulgarian flora. However, there is still much to be studied about the phytochemical and biological characteristics of the species. The present study aimed to determine the chemical composition of the ethanol extracts of aerial plant parts, by GC-MS analysis, and to thereby evaluate their in vitro antitumor and antibacterial properties. All three extracts were tested against the HT-29 and PC3 tumor cell lines using the MTT assay. Fifty-six components were identified from leaf, flower, and stem bark extracts, and over 10% were the following constituents: pyrogallol, α-terpinyl acetate, neryl acetate, and α-terpinyl isobutanoate. The oxygenated monoterpenes predominated in the extracts, followed by the oxygenated aliphatics and phenylpropanoids. Significant antiproliferative activity on the HT-29 cell line (IC50-21.44 µg/mL and 23.63 µg/mL, respectively) was found for the flower and leaf extracts. Antibacterial activity was established for the following bacteria strains: Bacillus subtilis ATCC 6633, Bacillus cereus NCTC 10320, Escherichia coli ATCC 8739, Pseudomonas aeruginosa ATCC 6027, and Proteus vulgaris ATCC 6380. The stem bark and flower extracts showed better antimicrobial potential. K. paniculata could be considered as a potential source of biologically active substances with antitumor and antibacterial properties.

Can Biomarkers Respond Upon Freshwater Pollution?-A Moss-Bag Approach.

Moss-bags were applied to study the effect of contamination in three standing water bodies in Bulgaria (Kardzhali, Studen Kladenets and Zhrebchevo Reservoirs), the first two with old industrial contamination and the last polluted with short-chain chlorinated paraffins (SCCPs). Fontinalis antipyretica Hedw. collected from background (unpolluted) site was placed in cages for a period of 30 days. The present study examined whether inorganic and organic pollution detected with moss-bags resulted in corresponding differences in molecular, chemical and micromorphological markers. Suppressed large subunit of ribulose-1,5-bisphosphate carboxylase (rbcL) expression was assessed in moss-bags from two of the reservoirs, contaminated with heavy metals. There was a decrease of the total phenolic content (TPC) in the moss-bags, which provides a basis for further studies of the chemical content of aquatic mosses. Fontinalis antipyretica also showed a response through leaf micromorphological characteristics. In the all three reservoirs, an increase of the twig leaf cell number was recorded (p ≤ 0.01 for Kardzhali and p ≤ 0.001 for Studen Kladenets and Zhrebchevo reservoirs), as well as of the stem leaf cell number in Zhrebchevo Reservoir (p ≤ 0.001). On the contrary, the width of the cells decreased in the studied anthropogenically impacted reservoirs. All three studied groups of biomarkers (molecular, chemical and micromorphological) appeared to be sensitive to freshwater pollution. The results achieved indicated that rbcL gene expression, TPC, cell number and size are promising biomonitoring tools.

Phenolic Profile and In vitro Antioxidant Activity of Endemic Bulgarian Carduus Species.

BACKGROUND: Plant species from genus Carduus are widely distributed in the world and represented in Bulgaria by 14 species. Previous investigations on this genus demonstrated a strong antioxidant potential of extract from some Bulgarian Carduus species. OBJECTIVE: The present study investigates the phenolic profile and the antioxidant potential of different extracts obtained from four endemic Compositae herbs, growing wild in Bulgaria: Carduus armatus Boiss and Heldr., Carduus candicans Waldst. et Kit ssp. globifer (Velen.) Kazmi., Carduus rhodopaeus Velen. and Carduus thracicus (Velen.) Hayek. MATERIALS AND METHODS: Antioxidant capacity of the obtained extracts was estimated with 2,2-diphenyl-1-picrylhydrazyl, 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid), and ferric reducing antioxidant power and copper reduction antioxidant assays. Phenolic profile was estimated by high performance liquid chromatography. RESULTS: Eleven phenolic acids and eight flavonoids were quantified in the inflorescences. Sinapic (2760.72 ± 15.68 µg/g dry weight [dw]), chlorogenic (2564.50 ± 19.73 µg/g dw) and ferulic acids (1648.71 ± 19.57 µg/g dw), as well as luteolin (2345.45 ± 18.61 µg/g dw) and apigenin (1332.75 ± 12.05 µg/g dw) were found to be the predominant compounds. The above contents are the highest values found in C. candicans ssp. globifer. The highest established antioxidant activity (AOA) was in favor of the ethanolic extracts, and the extract of C. rhodopaeus affirmed with the highest AOA among the investigated plant species. CONCLUSION: All identified phenolic compounds were reported for the 1(st) time in the studied endemic Carduus species, as well as their antioxidant capacities. The present study revealed that these plant species could be used as sources of antioxidants with potential medicinal properties. SUMMARY: Phenolic acids and flavonoid profiles of four endemic compositae herbs, growing wild in Bulgaria: Carduus armatus, Carduus candicans ssp. globifer, Carduus rhodopaeus and Carduus thracicus, were quantified for the first time by high performance liquid chromatography (HPLC). Eleven phenolic acids and eight flavonoids were determined in the inflorescences. Sinapic, chlorogenic and ferulic acids, as well as luteolin and apigenin were found to be the predominant compounds. The highest values of established phenolic compounds were found in C. candicans ssp. globifer. The studied plant extracts of Carduus species possessed antioxidant activity in favor of C. rhodopaeus and results confirmed 70 % ethanol as more appropriate solvent. The present study revealed that these plant species could be used as sources of antioxidants with potential medicinal properties.