Goran Anačkov

Plantain (Plantago L.) Species as Novel Sources of Flavonoid Antioxidants

To examine the antioxidant properties of methanol extracts of selected Plantago species (P. argentea Chaix., P. holosteum Scop., P. major L., P. maritima L., and P. media L.), various assays that measure free radical scavenging ability were carried out: DPPH, hydroxyl radical, superoxide anion, and nitric oxide scavenger capacity tests, reducing power (FRAP) assay, and Fe(2+)/ascorbate induced lipid peroxidation. In all of the tests extracts showed a potent antioxidant effect compared with BHT, a well-known synthetic antioxidant, and the extract of P. major, accepted as an official remedy. Besides, in examined extracts the total phenolic amount (ranging from 38.43 to 70.97 mg of GAE/g of dw) and the total flavonoid content (5.31-13.10 mg of QE/g of dw) were determined. Furthermore, the presence and content of selected flavonoids (luteolin-7-O-glucoside, apigenin-7-O-glucoside, luteolin, apigenin, rutin, and quercetin) were studied using LC-MS/MS technique. LC-MS/MS analysis showed noticeable qualitative and quantitative differences between the species according to which the examined Plantago species could be regarded as a possible new source of natural antioxidants. In this study three of the species examined, P. maritima, P. argentea, and P. holosteum, have been analyzed for the first time.

Exploring Equisetum arvense L., Equisetum ramosissimum L. and Equisetum telmateia L. as sources of natural antioxidants.

The antioxidant and scavenging activities of above ground parts of Equisetum arvense L., Equisetum ramosissimum L. and Equisetum telmateia L. phosphate buffer (pH 7) extracts were investigated. Activities of antioxidant enzymes (superoxide dismutase, catalase, guaiacol peroxidase and glutathione peroxidase), quantities of reduced glutathione, malonyldialdehyde, superoxide and hydroxyl radicals and flavonoid, soluble protein, chlorophyll a, b and carotenoid contents were determined. The total antioxidant capacity was determined by ferric reducing antioxidant power (FRAP) assay. The Equisetum telmateia extract demonstrated scavenging and antioxidant properties better than Equisetum ramosissimum and Equisetum arvense. The ESR signal of DMPO‐OH radical adducts in the presence of Equisetum telmateia phosphate buffer (pH 7) extract was reduced by 98.9% indicating that Equisetum telmateia could be a useful source of antioxidants with huge scavenging ability. Copyright

Chemical Composition of Essential Oil and Leaf Anatomy of Salvia bertolonii Vis. and Salvia pratensis L. (Sect. Plethiosphace, Lamiaceae)

The taxonomical relationship between Salvia pratensis and S. bertolonii has been unclear for a long time. Salvia bertolonii has alternatively been considered a synonym, a subspecies, a problematic subspecies and a form of Salvia pratensis. However, both these two species are sometimes used in traditional medicine instead of sage (Salvia officinalis) or as an adulteration for the same drug. In order to confirm the status of S. bertolonii, together with the potential identification characteristics for differentiation from sage, both taxa were analyzed through the analysis of their essential oils, together with the micromorphological characteristics of the leaf surface and the anatomy and morphology of the leaves. The obtained results show that there are clear differences in the quantity of essential oil (0.073% for S. pratensis and 0.0016% for S. berolonii). The major compound in the essential oil of S. pratensis was E-caryophyllene (26.4%) while in S. berolonii essential oil caryophyllene oxide was the major component (35.1%). The micromorphological differences are also pronounced in the leaf indumentum (density and distribution of certain types of non-glandular and glandular trichomes). Clear distinction between the investigated Salvia species is also observed in the leaf anatomy (in S. pratensis leaves are thinner, palisade tissue is made of 1-2 layers of cells, and leaves of S. bertolonii are characterized by 2-3 layers of palisade tissue cells, and consequently thicker).

Structural investigations of trichomes and essential oil composition of Salvia verticillata

Abstract.Krstic L., Malencic Dj. and Anackov G. 2006. Structural investigations of trichomes and the essential oil composition of Salvia verticillata. Bot. Helv. 116: 159 – 168.The morphology of glandular and non-glandular trichomes and the essential oil composition of Lamiaceae play an important role in the ecology of these species as well as for their industrial use; they may also serve as taxonomic criteria. We studied the trichome anatomy and essential oil composition of three wild-growing populations of Salvia verticillata in Serbia to determine how strongly these traits vary among and within Salvia species, and to evaluate their potential taxonomic or economic significance. Microscopic investigations of leaf and calyx indumentum revealed that S. verticillata has the same types of peltate and capitate trichomes as other Salvia species. In combination, however, the trichome characteristics of S. verticillata differed from previously examined Salvia species and might therefore help in species identification. The density of glandular trichomes differed among the three populations and was lowest in a dry steppe habitat, but trichome density also varied substantially among individual leaves within each population. The essential oil content (determined gravimetrically in n-hexane) ranged from 0.40% to 0.42% of dry mass. In total, 39 different oil components were identified using GC-MS. Oil composition varied considerably among the three populations. Only three of the 39 compounds were well represented (>0.1% of oil) in all three populations: The main component was germacrene D in two populations (48.0% and 24.6% of oil, respectively) but (E)- caryophyllene (10.2%) in the third. Low oil content and high variability in oil composition may restrict the industrial use of wild-growing S. verticillata plants.

Impact of origin and biological source on chemical composition, anticholinesterase and antioxidant properties of some St. John's wort species (Hypericum spp., Hypericaceae) from the Central Balkans.

The study shows the influence of the origin of plant material and biological source on the in vitro antioxidant (neutralization of DPPH and OH radical, nitric oxide, and inhibition of lipid peroxidation) and anticholinesterase activity of chemically characterized and quantified ethanol extracts of ten St. John’s wort samples. The investigated samples were: five Hypericum perforatum species representatives collected at different localities, one commercial sample of Hyperici herba purchased at a local market and four Hypericum species autochtonous to the Balkan Peninsula (H. maculatum subsp. immaculatum, H. olympicum, H. richeri subsp. grisebachii and H. barbatum). All the examined extracts exhibited notable antioxidant potential, but in most of the cases indigenous Hypericum species expressed stronger effects compared to the original source of the drug, H. perforatum. The changes in the content of phenolic compounds, especially flavonoids, hyperforin and hypericin, related to the source of the drug affected the investigated activities. Since all of the investigated species have shown prominent inhibition of acetylcholinesterase in vitro activity, they could be further investigated as potential substances in preventing of Alzheimer’s disease.

Comparative study of biological activities and phytochemical composition of two rose hips and their preserves: Rosa canina L. and Rosa arvensis Huds.

The aim of this study was to compare phenolic profile, vitamin C content, antioxidant, anti-inflammatory and cytotoxic activity of rose hips and the preserves (purée and jam) of two Rosa species: renowned Rosa canina L. and unexplored Rosa arvensis Huds. The liquid chromatography-tandem mass spectrometry analysis of 45 phenolics resulted in quantification of 14 compounds, with quercitrin, gallic and protocatechuic acids as the most dominant. High antioxidant potential of R. canina and a moderate activity of R. arvensis extracts were determined through several assays. Purée of both species and methanol extract of air-dried R. canina hips showed some anti-inflammatory (cyclooxygenase-1 and 12-lipooxygense inhibition potency) activity. Purée of R. canina exerted cytotoxic activity only against the HeLa cell line among several others (HeLa, MCF7, HT-29 and MRC-5). The presented results support traditional use of rose hips and their fruit preserves as food with health and nutritional benefits.

The distribution and the abundance of hydrophytes along the Danube River in Serbia

The Danube River runs through Serbia with flow 588 km long. Different hydrological, geological and ecological conditions occuring along the Danube in Serbia enables its division into four sections: The upper region, Đerdap I accumulation, Đerdap II accumulation, and the lower region. Each section is characterized by unique plant species diversity, frequency of occurrence and distribution pattern. The species diversity ranged from low in the upper reach, to high or very high in the impoundments and decreased in the lower reach. This paper provides a basis for a detailed study on the influence of large power plant impoundments over the aquatic vegetation of rivers.

Contact Sensitivity in Patients with Psoriasis in Vojvodina

Background: Immunologic studies indicate that psoriasis may represent an organ-integrated response. Objective: We assessed contact hypersensitivity in plaque-type psoriasis, its frequency, etiology and association with severity of psoriasis. Methods: Contact hypersensitivity was defined as a positive patch test (PT) to at least one of 44 ubiquitous contact allergens. Patients with exclusively plaque psoriasis and control groups (patients with allergic contact or extrinsic atopic dermatitis and healthy persons) were tested with the European standard series; plant-related standard allergens; Compositae allergens, and our own extracts from Compositae plants ubiquitous in Vojvodina. Sensitization rates to allergens were standardized for age and sex, and rates in women and in men were both standardized for age. Disease severity was evaluated using the Psoriasis Area Severity Index score. Results: 15,123 PTs were performed. Among psoriatic patients in Vojvodina, the overall rate of sensitivity, standardized for age and sex, was 18.9%. Rates in women (27.7%) and men (5.8%), both standardized for age, were significantly different. Male psoriatic patients reacted less than healthy males, the difference being on the margin of significance. There was no relationship between severity of disease and PT reactivity. Yarrow extract, nickel and a Compositae mix were the most common allergens that produced positive reactions. Conclusion: Although patch testing in psoriatic patients can be quite challenging, time-consuming and difficult, it will provide further insight into the pathophysiology of psoriasis. Factors other than different exposure to allergens may also be responsible for a sex-related difference in contact sensitivity. Future studies should focus on this field.

Biochemical Characterization of Helichrysum italicum (Roth) G.Don subsp. italicum (Asteraceae) from Montenegro: Phytochemical Screening, Chemotaxonomy, and Antioxidant Properties

The chemical composition and antioxidant properties of the essential oil and EtOH extract of immortelle (Helichrysum italicum (Roth) G.Don subsp. italicum, Asteraceae) collected in Montenegro were evaluated. The essential oil was characterized by GC/MS analysis, and the content of total phenolics and flavonoids in the EtOH extract was determined using the FolinCiocalteu reagent. The free‐radical‐scavenging capacity (RSC) of both the essential oil and the EtOH extract was assessed with the 2,2‐diphenyl‐1‐pycrylhydrazyl (DPPH) method. Moreover, the inhibition of hydroxyl radical (.OH) generation by the EtOH extract of immortelle was evaluated for the first time here. Neryl acetate (28.2%) and γ‐curcumene (18.8%) were the main compounds in the essential oil, followed by neryl propionate (9.1%) and ar‐curcumene (8.3%). The chemical composition of the oils of the examined and additional 16 selected Helichrysum italicum taxa described in literature were compared using principal component (PCA) and cluster (CA) analyses. The results of the statistical analyses implied the occurrence of at least four different main and three subchemotypes of essential oils. Considering the antioxidant properties, the EtOH extract of immortelle exhibited similar potential as propyl gallate and quercetin, while the essential oil exhibited relatively weak DPPH.‐scavenging capacity.

Alien invasive neophytes of the Southeastern part of the Pannonian Plain

Floristic diversity in the Pannonian Plain is the result of mosaic habitat composition, characterized by steppes, saline meadows, and lowland forests along major rivers. In the last few centuries, the flora has become enriched by species introduced by human activities, such as colonization and industrialization. Presently it consists of nearly 1800 species. As a result of 30 years of field research as well as herbarium and literature searches, a list of invasive alien neophytes has been compiled and is presented here. The list includes 152 species, classified as either naturalized, casual, or unknown. One of the species is listed in Red Book of Serbian Flora. Eighty species are naturalized, and 61 are casual species. The majority of the introduced flora originates from North America (66) and the Old World (43). Most alien invasive species of the Southeastern part of the Pannonian Plain, were introduced accidentally by seed material or deliberately by nursery saplings. The annual and perennial herbaceous plants, which flower in the summer and summer-autumn periods, are represented in the greatest numbers. The alien flora was further analyzed with respect to taxonomy, life forms, pollination, and seed dispersal mode. The greatest number of alien species appears in anthropogenic and semi-natural habitats (59). In natural habitats, 45 invasive taxa were recorded, representing a conservation threat to biodiversity.