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In the present study, the surface and external structure of petals, sepals, hypanthium and pedicel of eight Rosa species were examined. Micromorphological investigation of distribution and dimension of different trichome types, which were found in analysed plant parts was done using scanning electron microscopy (SEM). Our results showed that the adaxial epidermal cells of all analysed petals formed papillae, whereas the abaxial epidermis was with flat walls and had cuticle striation. Two types of trichomes, the capitate glandular and nonglandular trichomes (ribbon and acicles) were observed. Ribbon trichomes were presented on the adaxial side of all analysed sepals with the exception of R. spinosissima samples. Multicellular glandular trichomes were distributed on the surface of the hypanthium and the sepals of R. gallica, R. micrantha, R. pendulina, R. tomentosa and R. villosa samples. In addition, they were presented on the pedicel of most analysed species, except in R. dumetorum samples. Micromorphological analysis had shown that capitate glandular trichomes, based on their size, can be classified into two typeslong and short trichomes. Their presence, type and length were specific for each analysed species. Additionally, acicles were found only on the pedicel and hypanthium of R. spinosissima and R. gallica samples. This study provided useful information of different trichomes of genus Rosa which could be helpful in more accurate plant determination.

Pear ( Pyrus communis L.) is consumed as fresh fruit, in numerous food products, and also used as a traditional remedy in various countries, including Serbia. In search of bioactive compounds, six traditional pear varieties (‘Vidovača’, ‘Lubeničarka’, ‘Karamanka’, ‘Jeribasma’, ‘Lončara’, ‘Takiša’) and wild pear from Serbia were investigated and compared with a commercial variety (‘Williams Bartlett’). The aim of this study was to determine the total phenolic and flavonoid contents, phenolic composition, antioxidant capacity, and antineurodegenerative activities of methanolic extracts of peel, flesh, and mixed peel and flesh of pear fruits. Phenolic composition of extracts was determined with HPLC-DAD, while the antioxidant activity of extracts was evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), and ferric-reducing antioxidant power (FRAP) assays. Bearing in mind that oxidative stress is closely linked to neurodegeneration, the antineurodegenerative potential of the extracts was assessed by the inhibition of acetycholineserase (AChE) and tyrosinase (TYR) activities. The extracts of traditional varieties, particularly peel extracts, had a high content of phenolics, as well as significant antioxidant and moderate antineurodegenerative potential, compared to the commercial variety. The highest contents of total flavonoids and individual compounds, such as arbutin and chlorogenic acid, as well as the strongest antioxidant and TYR inhibitory activities were reported for the ‘Takiša’ peel extract. The performed analyses have revealed that fruits of traditional Serbian pear varieties are rich in bioactive components and could be used as functional food and for possible nutraceutical applications, to prevent diseases induced by oxidative stress.

In the most recent taxonomic literature, Centaurea crnogorica and C. gjurasinii, two neglected yellow-flowered taxa of C. sect. Acrocentron, have been treated as heterotypic synonyms of C. chrysolepis. Based on morphometric study, and study of type specimens and living plants from the locus classicus of C. gjurasinii and the only known population of C. crnogorica, as well as from several populations of C. chrysolepis from Eastern Serbia, we have found morphological differences among these three taxa. This has allowed us to re-evaluate taxonomic status of both, C. crnogorica and C. gjurasinii, which merit the species rank. The names C. crnogorica and C. gjurasinii are typified, and new expanded descriptions and illustrations are provided. We assessed also their conservation status. Because of their rarity, both taxa being narow endemics of two mountain ranges in Montenegro (the Durmitor Mts. and the Prokletje Mts.) and having small population sizes, C. crnogorica and C. gjurasinii are evaluated as critically endangered at national and global levels.

The main objective of this research was to evaluate the impact of methanolic, ethanolic and aqueous extracts of Origanum majorana L., Origanum vulgare L., Teucrium chamaedrys L., Teucrium montanum L., Thymus serpyllum L. and Thymus vulgaris L. (Lamiaceae) on the effects of free radicals using different model systems. The extracts were characterized on the basis of the contents of total phenolics, phenolic acids, flavonoids and flavonols, and also using high-performance liquid chromatography with diode-array detection. Antioxidant activity in vitro was assessed using DPPH assay. The genoprotective properties were tested using plasmid relaxation assay on pUC19 E. coli XL1-Blue, while SOS/umuC assay on Salmonella typhimurium TA1535/pSK1002 and Comet assay on human lung fibroblasts were used to assess the antigenotoxicity of the extracts. Ethanolic extracts had the most phenolics (up to 236.20 mg GAE/g at 0.5 mg/mL), flavonoids (up to 42.47 mg QE/g at 0.5 mg/mL) and flavonols (up to 16.56 mg QE/g at 0.5 mg/mL), and they exhibited the highest DPPH activity (up to 92.16% at 0.25 mg/mL). Interestingly enough, aqueous extracts provided the best protection of plasmid DNA (the lowest IC50 value was 0.17 mg/mL). Methanolic extracts, on the other hand, most efficiently protected the prokaryotic DNA, while all the extracts had a significant impact against genomic damages inflicted on human fibroblasts. O. vulgare extracts are considered to be the most promising in preserving the overall DNA integrity against oxidative genomic damages. Moreover, HPLC-DAD analysis highlighted rosmarinic acid as the most abundant in the investigated samples (551.45 mg/mL in total in all the extracts), followed by luteolin-7-O-glucoside (150.19 mg/mL in total), while their presence correlates with most of the displayed activities. The novelty of this study is reflected in the application of a prokaryotic model for testing the antigenotoxic effects of Lamiaceae species, as no previous reports have yet been published on the genoprotective potential of these species.

Juniperus communis or common juniper is the only conifer with pan-arctic distribution due to its polymorphic nature and high phenotypic plasticity. This taxon also produces large amounts of essential oil, which are stored in resin ducts in leaves (needles) and seed-cones. Leaf terpenoid composition of 97 individuals of common juniper from five populations in north-western Balkans was investigated. In total, 135 compounds were identified, representing on average 99.2% of the composition. Sabinene was the most abundant compound in most of the samples, followed by α- pinene and terpinen-4-ol. Two populations that were located deeper into the peninsula had a higher fraction of sesquiterpenes in comparison to the more coastal populations and one Julian Alps population. Multivariate statistical analyses also confirmed the separation of populations based on the essential oil composition. There was no correlation found between chemical composition and environmental factors. Chemophenetic implications are briefly discussed.

Spices are not only just herbs used in culinary for improving the taste of dishes, they are also sources of a numerous bioactive compounds significantly beneficial for health. They have been used since ancient times because of their antimicrobial, anti-inflammatory and carminative properties. Several scientific studies have suggested their protective role against chronic diseases. In fact, their active compounds may help in arthritis, neurodegenerative disorders (Alzheimer’s, Parkinson, Huntington’s disease, amyotrophic lateral sclerosis, etc.), diabetes, sore muscles, gastrointestinal problems and many more. In the present study, possible roles of spices and their active components, in chronic diseases (cancer, arthritis, cardiovascular diseases, etc.) along with their mechanism of action have been reviewed.

Pinus mugo Turra sensu stricto (dwarf mountain pine) is a species native to central and southern Europe, which performs a key role in subalpine areas by preventing avalanches and soil erosion. In t...

In this study, we examined Salvia amplexicaulis Lam., S. jurisicii Košanin and S. ringens Sibth. & Sm. collected in the Republic of North Macedonia, which are for the first time subjected to detailed micromorphological and structural analysis using light and scanning electron microscopy. The nutlets and mucilage were additionally subjected to spectroscopic analysis using Raman and Attenuated Total Reflectance Fourier Transform Infrared spectroscopy. The anatomical structure of stems and leaves is described and compared. The stems, leaves and calyces bear numerous one- and multi-cellular nonglandular trichomes, and various peltate, capitate and digitiform glandular trichomes. The nutlets differ in size and shape, as well as in myxocarpy. The nutlets predominantly contained α-linolenic and linoleic acid, whereas the mucilages are primarily formed of polysaccharides. The results obtained in this study confirmed the importance of micromorphological and anatomical analysis of Salvia L. spp. plant parts, particularly trichomes and nutlets, and hence contributing to the knowledge about the variety of micromorphological characteristics within the genus Salvia L. This is the peer-reviewed version of the article: Alimpić-Aradski, A.; Janosević, D.; Pećinar, I.; Budimir, S.; Dajić-Stevanović, Z.; Matevski, V.; Marin, P. D.; Duletić-Lausević, S. Micromorphological and Anatomical Characteristics of Salvia Amplexicaulis Lam., S. Jurisicii Kosanin and S. Ringens Sibth. & Sm. (Lamiaceae). Plant Biosystems 2021, 155 (1). [https://doi.org/10.1080/11263504.2020.1727976].

Paper description: Lamium purpureum L. (Lamiaceae) glandular trichomes and the composition of essential oil were analyzed. Two types of trichomes, non-glandular and glandular (peltate and capitate) were identified. The main compounds of essential oil were trans -anethole (59.61%), fenchone (6.54%), α -pinene (6.14%) and β -pinene (4.98%). The antioxidant activities of the ethanolic plant extracts were determined using DPPH, ABTS, FRAP and TRP assays. Antifungal activity of the ethanolic extract was observed, While the extracts did not exhibit fungicidal activity, morphophysiological changes (sporulation loss, depigmentation of conidial heads) were observed in all tested isolates. L. purpureum could be a new potential source of natural antioxidants. Abstract : The aim of this study was to investigate the structure of glandular trichomes and essential oil composition of Lamium purpureum L. (Lamiaceae) and to determine potential biological activities of its different extracts. The micromorphology and distribution of trichomes on vegetative plant parts were investigated using scanning electron microscopy (SEM). Two types of trichomes were identified: non-glandular (simple, uniseriate, multicellular) and glandular (both peltate and capitate). The essential oil of L. purpureum was isolated by hydrodistillation and analyzed by gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS). The main compounds were trans -anethole (59.61%), fenchone (6.54%), α -pinene (6.14%) and β -pinene (4.98%). The total phenol and flavonoid contents of 10%, 30% and 50% ethanol extracts were determined spectrophotometrically. The free radical scavenging and reducing power activity of extracts were analyzed using the DPPH and ABTS radical scavenging methods, and the ferric reducing antioxidant power (FRAP) and total reducing power (TRP) assays. The 50% ethanolic extract exhibited the strongest antioxidative activity while the lowest was determined for the 10% ethanolic extract. Antifungal activity of L. purpureum hydroalcoholic extracts was evaluated against eight Aspergillus species by the microdilution method. Light microscopy (LM) demonstrated structural abnormalities of the tested Aspergilli . While the plant extracts did not exhibit fungicidal activity, morphophysiological changes (sporulation loss and depigmentation of conidial heads) were observed in all tested isolates. https://doi.org/10.2298/ABS200117019G Received: January 17, 2020; Revised: February 25; Accepted: April 7, 2020; Published online: April 23, 2020 How to cite this article: Grujic SM, Savkovic ŽD, Ristic MS, Džamic AM, Ljaljevic Grbic MV, Vukojevic JB, MarinPD. Glandular trichomes, essential oil composition, anti- Aspergillus and antioxidative activities of Lamium purpureum L. ethanolic extracts. Arch Biol Sci. 2020;72(2):253-63.

The content of nonacosan-10-ol and n-alkanes in needle waxes of twelve Pinus taxa (Pinus halepensis, P. heldreichii, P. mugo, P. nigra ssp. nigra, P. nigra ssp. dalmatica, P. peuce, P. pinaster, P. pinea, P. ponderosa, P. strobus, P. sylvestris, and P. wallichiana) were examined. In eight Pinus taxa (except of P. halepensis, P. pinaster, P. heldreichii and P. peuce) nonacosan-10-ol contents were examined for the first time. In all examined pines C29, C27 or C25 were the main n-alkane compounds. C range was mainly 18-33. In six Pinus taxa (P. mugo, P. nigra ssp. dalmatica, P. sylvestris, P. pinea, P. strobus and P. wallichiana) CPI and ACL values were examined for the first time. In the plane of Axis 1 (PCA), P. strobus, P. mugo, and P. wallichiana were strongly separated from P. heldreichii and P. pinea. In the plane of Axis 2, P. peuce, P. strobus, and P. wallichiana as well as P. heldreichii and P. pinea diverged from other examined species. In DA P. heldreichii, P. strobus, P. peuce, and P. wallichiana diversed. CA divided sections Pinus and Pinaster from section Strobi. This is the peer-reviewed version of the article: Nikolic, B. M.; Đorđević, I.; Todosijevic, M.; Mitic, Z.; Stefanović, M. A.; Stankovic, J. M.; Bojović, S.; Tešević, V.; Marin, P. D. Diversity of Nonacosan-10-Ol and n-Alkanes among 12 Pinus Taxa. Plant Biosystems - An International Journal Dealing with all Aspects of Plant Biology 2020, 0 (0), 1–8. [https://doi.org/10.1080/11263504.2020.1857867]

This study was designed to evaluate the genoprotective, antigenotoxic, as well as antitumor potential of methanolic, ethanolic, and aqueous extracts of Melissa officinalis, Mentha × piperita, Ocimum basilicum, Rosmarinus officinalis, Salvia officinalis, and Satureja montana (Lamiaceae), in different model systems. The polyphenols in these extracts were quantified both spectrophotometrically and using HPLC-DAD technique, while DPPH assay was used to assess the antioxidant activity. The genoprotective potential was tested on pUC19 Escherichia coli XL1-blue, and the antigenotoxicity on Salmonella typhimurium TA1535/pSK1002 and human lung fibroblasts, while the antitumor activity was assessed on colorectal cancer cells. Rosmarinic acid, quercetin, rutin, and luteolin-7-O-glucoside were among the identified compounds. Methanolic extracts had the best DPPH-scavenging and SOS-inducing activities, while ethanolic extracts exhibited the highest antigenotoxicity. Additionally, all extracts exhibited genoprotective potential on plasmid DNA. The antitumor effect was mediated by modulation of reactive oxygen species (ROS), nitric oxide (NO) production, and exhibition of genotoxic effects on tumor cells, especially with O. basilicum ethanolic extract. Generally, the investigated extracts were able to provide antioxidant protection for the acellular, prokaryotic, and normal human DNA, while also modulating the production of ROS and NO in tumor cells, leading to genotoxicity toward these cells and their decrease in proliferation.

Based on previous investigations where bis-bibenzyls isolated from liverworts showed various biological activities (cytotoxic, antimicrobial, and antiviral), we investigated their cytotoxic activity in several human cancer cell lines. From the methylene-chloride/methanol extract of the liverwort Pellia endiviifolia, three bis-bibenzyls of the perrottetin type were isolated, namely perrottetin E, 10′-hydroxyperrottetin E, and 10,10′-dihydroxyperrottetin E. The last two were found for the first time in this species. Their structures were resolved using 1D and 2D NMR, as well as by comparison with data in the literature. Cytotoxic activity of the isolated compounds was tested on three human leukemia cell lines, HL-60 (acute promyelocytic leukemia cells), U-937 (acute monocytic leukemia cells), and K-562 (human chronic myelogenous leukemia cells), as well as on human embryonal teratocarcinoma cell line (NT2/D1) and human glioblastoma cell lines A-172 and U-251, and compared to the previously isolated bis-bibenzyls (perrottetins) of similar structure. The isolated compounds exhibited modest activity against leukemia cells and significant activity against NT2/D1 and A-172. Overall, the most active cytotoxic compounds in this investigation were perrottetin E (1), isolated in this work from Pellia endiviifolia, and perrottetin F phenanthrene derivative (7), previously isolated from Lunularia cruciata and added for a comparison of their cytotoxic activity.

Satureja kitaibelii Wierzb. ex Heuff. is endemic species of central Balkan that lives along the Balkan mountains range. The aim of this research was to study the variability of n-alkane profiles of four S. kitaibelii populations growing wild in Serbia in regards to biogeography and chemotaxonomy. Using GC/MS and GC/FID, seventeen n-alkane homologues (C19–C35) were identified. n-Hentriacontane (C31) was dominant in all populations, except in population Djerdap and Rtanj were it was co-dominant with n-nonacosane (C29). Population Temsica and Poganovo also had high percentages of n-nonacosane and n-tritriacontane (C33). However, these two compounds had similar quantities in population Poganovo. Statistical analyses showed differences between studied populations based on n-alkane profiles. Moreover, geographically closer populations had a similar profile.

Paper description: There are no studies on the influence of environmental factors and their correlation with pine oleoresin composition from the leaves of several Balkan pines. Oleoresins were isolated from Bosnian and Macedonian pine collected from nine localities, including two sympatric. The obtained composition was analyzed statistically to assess the correlation with bioclimatic, geological and geographical parameters. Three chemotypes were present in Bosnian pine leaf oleoresins, while only one was found in Macedonian pine. While the oleoresin profiles showed strong species-dependent composition and variability, environmental factors, including geological substrate type and exposition must also be taken into account. Abstract: Many studies show the influence of the environment on terpene composition, but not many of them deal with the terpene composition variability in correlation with environmental factors in Pinus. We chose two endemic species – P. heldreichii Christ. (Bosnian pine) and P. peuce Griseb. (Macedonian pine) – two relict and (sub)endemic species of the Balkan peninsula. They mainly form pure stands but can appear in mixed populations, most commonly with each other. These species belong to different subgenera and thus differ greatly, especially in needle anatomy. α -pinene, limonene and germacrene D predominated in Bosnian pine leaf oleoresins, while Macedonian pine oleoresin contained high amounts of α -pinene. Furthermore, there were three chemotypes present in Bosnian, and only one in Macedonian pine. Oleoresins of Bosnian pine did not show correlation with climate, however, its composition changed in response to geological substrate type. Macedonian pine oleoresin showed a high correlation with the climate but changed only a little in response to geological substrate type. The oleoresin profiles showed strong species-dependent composition and variability. However, since each species expressed different responses to tested environmental conditions, it is important to take into consideration these variables when analyzing natural populations of the species. https://doi.org/10.2298/ABS190705045R Received: July 5, 2019; Revised: July 18, 2019; Accepted: July 24, 2019; Published online: August 1, 2019 How to cite this article: Rajcevic N, Nikolic B, Marin PD. Different responses to environmental factors in terpene composition of Pinus heldreichii and P. peuce : Ecological and chemotaxonomic considerations. Arch Biol Sci. 2019;71(4):629-37.

This research was aimed to assess the potential of Glechoma hederacea, Hyssopus officinalis, Lavandula angustifolia, Leonurus cardiaca, Marrubium vulgare and Sideritis scardica (Lamiaceae) methanolic, ethanolic and aqueous extracts against the damaging effects of oxidative stress using different experimental models. The chemical characterization was done spectrophotometrically by quantifying total phenolics, phenolic acids, flavonoids and flavonols in the extracts, as well as by employing HPLC-DAD technique. Moreover, DPPH assay was used to assess the extracts' radical scavenging potential. Genoprotective properties of the extracts were evaluated using plasmid pUC19 Escherichia coli XL1-Blue, whereas their antigenotoxic potential was determined using Salmonella typhimurium TA1535/pSK1002 and normal human lung fibroblasts. All of the extracts showed antioxidant activity in DPPH assay. Furthermore, the results have shown that aqueous extracts provided the best protection for plasmid DNA, while alcoholic extracts most effectively contributed to the preservation of prokaryotic DNA. Additionally, each of the tested samples significantly protected the eukaryotic cells against genomic damages. Finally, despite not showing exceptional results in DPPH assay, S. scardica extracts are regarded as the most favorable in maintaining the integrity of DNA, which might be due to high quantities of phenolics such as quercetin (up to 17.95 mg g-1), naringin (up to 5.07 mg g-1) and luteolin-7-O-glucoside (up to 3.54 mg g-1). Overall, this comprehensive concept highlights the ability of these Lamiaceae species to safeguard the DNA from reactive oxygen species, to curtail the inflicted damage and also improve the efficiency of the DNA repair mechanisms, while emphasizing the importance of polyphenols as their active principles.

The essential oil (EO) composition of Satureja plants is highly variable. Recent studies suggest that there is an even difference in the EO composition from different plant organs within the same plant. This study aims to examine the chemical profile of EOs and the micromorphological characteristics of different organs of three Satureja species. The relationship between the number of glandular trichomes and EOs profile and relative yield is also investigated. Individuals from five populations were visualized using a scanning electron microscope, while EOs of leaves, calyces, corollas, and whole aerial parts were isolated using simultaneous distillation and extraction and analyzed using gas chromatography/mass spectrometry. Three types of glandular trichomes were detected. Peltate trichomes were present on all plant organs of studied species, while two types of capitate trichomes show different organ and species preferences. The EOs profiles differed across the plant parts, but showed a species specific composition. Univariate and multivariate statistics were used to show a correlation between the peltate trichomes and EO yield, and chemophenetic significance of EO profiles.

Prunus spinosaleaf extracts in solvents of different polarity (water, ethanol and acetone), their phenol,flavonoid and anthocyanin contents and biological properties were the object of this study. The richest in phenols as well as in flavonoids was acetone extract with 181.19 mg GAE and 80.10 mg QE per gram of dry extract, respectively. Moreover, the quantity of anthocyanins obtained byHPLC analysis was also the highestin acetone sample. Examined samples possessed antioxidant properties evaluated through fourin vitroassays(DPPH, ABTS, FRAP and TRC). The acetone extract wasproved to be the best antioxidant among testedsamples, which could be ascribed to polyphenols, especially anthocyanins. The aqueous and the ethanolextractexhibited antibacterial effects, being particularlyactive againstB. cereusandE. cloacaeT. viride P.funiculosum,P. ochrochloronP. verrucosumvar.cyclopiumwere the most susceptible among fungalmicroorganisms examined. Both, the aqueous and theethanol extract expressed inhibitory activity towardsenzymes linked to diabetes mellitus type II. Additionally, the ethanol extract showedsignificantly higherpotential in inhibitingα-glucosidase than the drug used as the positive control. Furthermore, the aqueoussample revealed antitumor effects on following malignant cell lines: HeLa, K562 and MDA-MB-453. Theresults presented herein suggest thatP. spinosaleaves should be considered as a natural source ofbioactivecompounds with potential application in phytopharmacy and food industry.

Lamiaceae (Labiatae) are an important group of medicinal plants, which have been used for treating heart disease in traditional medicine for centuries. These mainly aromatic plants are used as essential oils, extracts or isolated components (polyphenols, phenolic compounds, terpenes, iridoids etc.). Some Labiatae species (more than 30, such as cornmint, lavender, patchouli, rosemary etc.) are famous for their use in essential oil production worldwide. In this review, cardioprotective effects of Lamiaceae and their active secondary metabolites, as well as mechanism of action against cardiovascular diseases (hypertension, angina pectoris, hyperlipidemia, thromboembolism, coronary heart disease, heart failure, venous insufficiency, arrhythmia) will be discussed. Use of Labiatae as food or food additives (such as spices) may prevent risk of cardiovascular diseases, diabetes and cancer. This approach is also described as a part of the article. Studies on developing new, effective and safe natural products from Lamiaceae (rich source of flavonoids and other active compounds) are promising and may offer prevention and treatment for patients with coronary disease and other related diseases.

We examined essential oils (EOs) of Chamaecyparis lawsoniana and Thuja plicata (Cupressaceae), their antifeedant activity on Lymanthria dispar larvae and their antimicrobial activity. Studies of EOs showed that these two conifer species differed both in content of terpene classes and the major compounds. Total monoterpenes strongly dominated in T. plicata (96.4%), while C. lawsoniana was rich in both mono- and sesquiterpenes (40.8% and 30.3%, respectively) as well as in diterpenes (19.1%). The most dominant compounds of C. lawsoniana EO were: limonene (16.7%), oplopanonyl acetate (14.5%), beyerene (10.1%), and 13-epi-dolabradiene (6.7%). The dominant compound of T. plicata EOs was α-thujone (76.9%), followed by relatively small amounts of β- thujone (5.3%), sabinene (4.5%) and terpinene-4-ol (3.2%). The difference in EO compositions of the conifers was reflected on Lymantria dispar larvae performance. Larvae fed on the leaf discs treated by C. lawsoniana EO had a slight phagostimulatory effect at lower concentration shown by higher relative rate of food consumption and relative growth rate than the larvae in the control group. Contrastingly, leaf discs treated with EO of T. plicata EO had an antifeedant effect and lower relative consumption rate (RCR) and relative growth rate RGR than the larvae in the control group. Both tested EOs influenced substantially the colony growth of the subjected Phytophthora plurivora and P. quercina. Namely, 100% inhibitory effect was recorded at concentration of 0.1% in the case of C. lawsoniana EO, whereas of T. plicata colonies did not grow at 0.5% concentration. The implications of these findings and possibility of using the tested EOs in further experiments in vitro and in vivo are discussed.

In leaf cuticular wax of Pinus pinaster, content of nonacosan-10-ol is high (77.1% on average). n-Alkanes ranged from C18 to C35 with the most dominant C29 (24.8%). The carbon preference index (CPItotal) ranged from 3.1 to 5.6 (4.0 on average), while the average chain length (ACLtotal) ranged from 14.0 to 17.0 (14.8 on average). Long-chain n-alkanes ( n-C25-35) strongly dominated (80.1%) over middle-chain ( n-C21-24 = 18.9%) and short-chain ( n -C18-20 = 0.9%) n-alkanes.

This is the first report on population differentiation based on composition of three different compound classes ( n -alkanes, primary alcohols and diterpenes) in the epicuticular waxes of European black pine ( Pinus nigra ) from Balkan Peninsula and Southern Carpathians. The n -hexane extracts of 263 samples, originating from 15 native populations of four P. nigra subspecies ( nigra , dalmatica , banatica and pallasiana ), were investigated by GC-MS and GC-FID analyses. In all studied populations n -alkanes represent the most abundant compound group, while the contents of diterpenes and alcohols were lower and variable. Multivariate statistical analyses (Canonical Discriminant Analysis and Agglomerative Hierarchical Clustering) suggest existence of two clearly differentiated chemical entities with the basic difference in the content of primary alcohols (especially cis -9-octadecen-1-ol and 1-hexadecanol). The first group (low alcohol content) included all populations of ssp. nigra , ssp. dalmatica and ssp. banatica , as well as the two northernmost populations of ssp. pallasiana from Serbia and Bulgaria, while the second group (high alcohol content) comprised individuals from Greek and Macedonian populations of ssp. pallasiana . According to the recently proposed concept of ancient split of major genetic lineages of P. nigra , both chemical entities detected in this study belong to the same (Balkan Peninsula) genetic group. Taxonomic implications of given results are briefly discussed in relation with previous data based on morphological and molecular characters.

Ethnopharmacological relevance River and mountain regions in Eastern and South-Eastern Serbia are geographically interesting and, historically they represent an important resource of plants used as food, spices and as remedies for treating many diseases. Different cultures have lived in these regions for ages. They have used wild plants and the methods of their preparation and application, which has remained throughout the history and now is passed on from generation to generation. The aim of the study is a survey of herbal drug uses for the specific ailment categories and their comparison between the two research regions. Methods Semi-structured anonymous ethnobotanical interviews were conducted. The interviews took place in the River Timok region and Mountain Svrljig region as they make two of the most interesting centers of plant biodiversity. Volunteers in the Timok region were 64 median age and in the Svrljig region - 73 median age. People were interviewed about the local names of plants, the preparation process and about which disease the plants were used for. Results and Discussion 161 Respondents from 10 Timok and 10 Svrljig municipality villages were interviewed and 2199 use-reports were recorded. The results of the ethnobotanical research showed 195 plant taxa from the Timok and Svrljig regions. In the Timok region, the recorded species were classified into 47 families and in the Svrljig region they were classified into 64 families. Out of 195 taxa used for medicinal purposes, only 52 species are also included in the European Pharmacopoeia 9.0. The most commonly used plants in Timok region were Hypericum spp., Matricaria chamomilla, Mentha x piperita, Urtica dioica, Juglans regia, while the residents of Svrljig region most frequently used Satureja montana, Sambucus nigra, Polygonum aviculare, Marrubium vulgare and Teucrium chamaedrys. Different statistical analyzes showed that Vlachs used more medical specimens per person than Serbs in the Timok region. The study demonstrated that female informants treated more body organs with medical species per person than male informants. There was a significant difference between the male and female respondents of the Svrljig region in terms of organ systems for which they used certain plant species. Conclusion The ethnopharmacological study showed a great importance of medicinal plants in the daily life of local communities. According to the analysis, it can be concluded that the village population of the Timok and Svrljig regions use medicinal plants to treat digestive tract problems rather than seeking professional medical attention in health facilities. The aerial part and rhizome of Elymus repens are used for digestive problems in both the Timok and Svrljig regions, and the use of this plant in for the treatment of digestive tract problems is not mentioned in the similar studies conducted in the Balkan region. Also, it can be observed that the population of the two different regions mainly use different herbal drugs to treat the same systems. The reason for that is the availability of certain plants that grow in the two different areas - river and mountain area.

This study was aimed to analyze the chemical composition and biological activities of extracts of Salvia fruticosa and S. lanigera originated from Libya. Dichloromethane, ethyl acetate, methanol, ethanol and water extracts obtained from wild growing plants were analyzed for the composition using HPLC-DAD, which revealed presence of phenolic acids and flavonoids, mainly in alcoholic and water extracts. Total phenolic and flavonoid contents, as well as anti-oxidant activity using 2.2-diphenyl-1-picrylhydrazyl (DPPH), 2.2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid diammonium salt (ABTS), ferric reducing anti-oxidant potential (FRAP) and β-carotene bleaching (β-CB) assays were determined spectrophotometrically. Extracts of S. fruticosa exhibited stronger activity in all applied assays, especially ethanol extract in DPPH assay (IC50, 29.55 μg/mL) and in β-CB assay (85.11%). The ethanol and water extracts were selected for further investigation, because of their wide usage in phytotherapy. The extracts were screened for the antimicrobial activity against 11 bacteria (six Gram-negative and five Gram-positive) and seven fungi using microdillution method. Ethanol extracts showed stronger activity than water extracts, particularly against Gram-positive bacteria. Trichophyton mentagrophytes was the most sensitive to the water extract of S. lanigera (MIC, minimal inhibitory concentration and MFC, minimal fungicidal concentration, 8 mg/mL). Cytotoxic activity on human carcinoma cell line HCT-116 was determined by 3-(4.5-dimethylthiazol-2-yl)-2.5-diphenyl tetrazolium bromide (MTT) cell viability assay, where only ethanol extract of S. fruticosa demonstrated certain activity (IC50, 375.96 μg/mL). The testing of anti-neurodegenerative activities of extracts showed better tyrosinase inhibiting effect (55.26–74.66%) than standard kojic acid (33.93–51.81%), while the extracts were less effective against acetylcholinesterase compared with standard galanthamine. According to the obtained results, S. fruticosa and S. lanigera originated from Libya have proved to be the promising source of natural compounds possessing a range of biological activities.

Many Eryngium species have been traditionally used as ornamental, edible or medicinal plants. The gas chromatography-flame ionization detector (GC-FID) and gas chromatography-mass spectrometry (GC–MS) analyses have shown that the major compounds in the aerial parts were spathulenol (in E. campestre and E. palmatum oils) and germacrene D (in E. amethystinum oil). The main compounds in the root oil were nonanoic acid, 2,3,4-trimethylbenzaldehyde and octanoic acid for E. campestre, E. amethystinum and E. palmatum, respectively. All the oils expressed the highest potential against Gram-positive bacteria Staphylococcus aureus as well as Gram-negative Klebsiella pneumoniae and Proteus mirabilis. Molecular docking analysis was used for determining a potential antibacterial activity mechanism of compounds present in the essential oils. Molecular docking confirmed that the binding affinity of spathulenol to the active site of tyrosyl-tRNA synthetase was the highest among the tested dominant compounds. Regarding the total phenolic content (determined by the Folin–Ciocalteu assay) and flavonoid content (evaluated using aluminum nitrate nonahydrate), the highest amount was found in the ethyl acetate extract of E. palmatum. The results of DPPH and ABTS assay indicated that the highest antioxidant activity was present in the water extract of E. amethystinum. Extracts of the aerial parts presented as minimum inhibitory concentration (MIC) expressed the activity in the range 0.004–20.00 mg/mL, with the highest activity exhibited by the acetone and ethyl acetate extracts against Proteus mirabilis. The obtained results suggest that Eryngium species may be considered a beneficial native source of the compounds with antioxidant and antimicrobial properties.

Comparative analysis of the chemical composition and biological activity of the essential oils from the needles of four Pinus species (P. mugo subsp. mugo, P. nigra subsp. nigra, P. sylvestris and P. peuce) from the central Balkans was performed. Although different chemical profiles of the dominant terpenes for each species were determined, essential oils were, generally, dominated by monoterpenes with α-pinene as one of the first two major volatiles. Tested oils showed inhibitory action against respiratory pathogenic bacterial strains, isolated from human swabs, in the range of 1.25–20.00 mg/mL (MICs) and bactericidal effect (MBCs) at concentrations from 2.50 to 40.00 mg/mL. Considering cell wall composition, there were no significant differences in sensitivity between Gram-positive and Gram-negative strains. In addition, the place of isolation did not provide evidences for higher susceptibility/resistance of the strains obtained from the particular infection site. On the other hand, toxicity of essential oils towards developing Drosophila melanogaster larvae was low, except P. mugo subsp. mugo oil, which did not show any signs of toxicity at concentrations up to 3%. Highest mortality of larvae and adults of D. melanogaster was noted for P. sylvestris essential oil with an estimated LC50 at the end of experiment (14 days) of 2.78% with a 95% confidence interval of 2.15–3.42%. Short-term 96 h LC50 values due to low mortality at larval stage could not be calculated for any oil and are expected to be much higher than 3%. According to the obtained antimicrobial activity and toxicity towards D. melanogaster, the oils can be arranged in the following order: P. sylvestris > P. peuce > P. nigra subsp. nigra > P. mugo subsp. mugo.

We examined for the first time the leaf micromorphology, phytochemistry and biological activity of the rare and stenoendemic Amphoricarpos elegans Albov (Compositae) from Georgia. Scanning electron microscopy (SEM) revealed the presence of glandular trichomes on the leaves, which appeared as glandular dots that are considered the main sites of biosynthesis and accumulation of sesquiterpene lactones. Using high-performance liquid chromatography (HPLC) and nuclear magnetic resonance (NMR) spectroscopy analyses, we identify and characterized 3-deoxyamphoricarpolide, a known sesquiterpene lactone for the genus Amphoricarpos Vis. Regarding chemotaxonomic significance, 3-deoxyamphoricarpolide represents a link between Balkan and Caucasian species of the genus. The antioxidative capacity of different leaf extracts, obtained using a Soxhlet extractor, was evaluated by two radical scavenging assays: DPPH (1,1-diphenyl-2-picrylhydrazyl) radical and the 2,2΄-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS), and ferric ion reducing antioxidant power (FRAP). The total phenolic and flavonoid contents were also determined. The highest antioxidative activity and the highest phenolic and flavonoid contents were detected in the methanol fraction, as a result of the contribution of not only phenols, but probably also lactones. The considerable antioxidative potential indicates possible applications in pharmacy and medicine., Financial support was provided by the Ministry of Education, Science and Technological Development of the Republic of Serbia (Project Nos. 173029 and 172053).