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Composition and antimicrobial activity of root, leaf, stem, flower and fruit essential oils from cultivated Pastinaca sativa subsp. sativa, and its two wild-growing relatives P. sativa subsp. urens and P. hirsuta (Apiaceae) were investigated. Twenty-nine hydrodistilled essential oils of plants from different localities and/or years were analysed by GC-FID and GC/MS. Dominant in root oils was myristicin (P. sativa) or apiole (P. hirsuta), in leaf and stem oils myristicin (cultivated plants) or γ-palmitolactone (wild-growing plants) and in flower and fruit oils aliphatic esters. Multivariate statistics (PCA, nMDS, UPGMA clustering) generally revealed separation of oils of investigated Pastinaca taxa and demonstrated their chemosystematic significance. One oil per each organ of all three plants (fifteen in total) was tested using microdilution method for activity against Candida tropicalis, C. parapsilosis, C. krusei, C. glabrata, C. albicans, Staphylococcus aureus, Bacillus cereus, Listeria monocytogenes, Escherichia coli, Salmonella Typhimurium and Enterobacter cloacae; MIC = 0.25–8 mg/mL, MBC(MFC) = 0.5–16 mg/mL.

This work was aimed at investigating the anatomy and composition of the essential oils from different organs of two Pimpinella species, P. tragium and P. saxifraga, originating from the central part of the Balkan Peninsula. The results of the analysis of the anatomy of the different organs of P. tragium and P. saxifraga showed that they were very similar in terms of the structure of the roots, stems and petioles. The roots showed a secondary structure and the stems had a primary structure with closed collateral vascular bundles. The petioles were cordatum, ribbed with an adaxial groove, and sparsely covered with short unior bicellular non-glandular trichomes with a pointed top (both species) or with long uniseriate, multicellular, non-glandular trichomes (P. saxifraga). Differences were observed in the anatomy of the leaves and in the indumentum of the fruits. The leaves of P. tragium were isobilateral, while those of P. saxifraga were dorsiventral. The P. tragium fruits were pubescent, covered with numerous, multicellular, uniseriate, straight or curved, gradually acuminate non-glandular trichomes, whereas the fruits of P. saxifraga were glabrous. These differences are of ecological and taxonomic significance. Secretory canals were observed in all the investigated organs of both species. The essential oils, isolated by hydrodistillation from the aerial parts with inflorescences, the roots from the flowering and fruiting period, and the fruits of both species, were analysed by GC-FID/MS. Although P. tragium and P. saxifraga essential oils showed certain differences in qualitative and quantitative patterns, all the oils were characterised by the presence of pseudoisoeugenol type phenylpropanoid epoxy-pseudoisoeugenyl-2-methylbutyrate, as well as by that of trinorsesquiterpenes pregeijerene, geijerene and trinoranastreptene. In addition, azulenes were present in all the root essential oils, whereas in the majority of the oils from the fruits and flowering aerial parts the common compound was the sesquiterpene β-bisabolene. The occurrence of phenylpropanoids of pseudoisoeugenol type and geijerenes is important from the chemotaxonomic point of view and confirms them as chemical markers of the Pimpinella species analysed so far. Cilj ovog istraživanja bio je analiza anatomske građe i hemijskog sastava etarskih ulja različitih organa dve vrste roda Pimpinella, P. tragium i P. saxifraga, poreklom iz centralnog dela Balkanskog poluostrva. Na osnovu rezultata analize unutrašnje građe različitih organa P. tragium i P. saxifraga može se zaključiti da postoje velike sličnosti u građi korena, stabla i lisne drške. Koren obe vrste bio je sekundarne građe, stablo primarne građe sa zatvorenim kolateralnim sprovodnim snopićima, a lisne drške srcolike, rebraste sa adaksijalnim žlebom, sporadično prekrivene kratkim jednoćelijskim ili dvoćelijskim, na vrhu zašiljenim nežlezdanim trihomama (obe vrste) ili dugim uniserijatnim, višećelijskim nežlezdanim trihomama (P. saxifraga). Uočene su razlike u anatomiji listova i in- dumentumu plodova ove dve vrste. Listovi P. tragium bili su izobilateralni, a P. saxifraga dorziventralni. Plodovi P. tragium bili su maljavi, prekriveni brojnim, višećelijskim, uniserijatnim, uspravnim ili zakrivljenim, postepeno zašiljenim nežlezdanim trihomama, dok su plodovi P. saxifraga bili goli. Ove razlike u anatomskoj građi su od ekološkog i taksonomskog značaja. U svim organima obe ispitivane vrste uočeno je prisustvo sekretornih kanala. Etarska ulja su izolovana postupkom destilacije vodenom parom iz različitih organa obe vrste, nadzemnog dela sa cvastima, korena prikupljenog u fazi cvetanja i u fazi plodonošenja i ploda i analizirana GC-FID/ MS metodom. Iako su ispitivana etarska ulja P. tragium i P. saxifraga bila donekle različitog kvalitativnog i kvantitativnog sastava, za sva etarska ulja bilo je karakteristično prisustvo fenilpropanoida pseudizoeugenol tipa epoksi-pseudoizoeugenil-2-metilbutirata, kao i trinorseskviterpena pregeijerena, geijerena i trinoranastreptena. Pored toga, u svim etarskim uljima dobijenim iz korena bili su prisutni azuleni, a u većini etarskih ulja plodova i nadzemnih delova u cvetu karakterističan sastojak bio je sekviterpen β-bisabolen. Prisustvo fenilpropanoida pseudoizoeugenol tipa i trinorsekviterpena tipa geijerena značajno je sa hemotaksonomskog aspekta i pot- vrđuje njihovu ulogu hemijskih markera do sada ispitivanih Pimpinella vrsta.

The importance of comparison in geography education is undoubtedly immense. Comparison as a process of thinking represents the oldest method for deriving geographical evidence and it is often used in teaching geography. It can be argued that this method is the most characteristic method for geography as a school subject. In addition to the development of the students’ ability to analyze, synthetize, generalize, and distinguish between the relevant and the irrelevant, this method can also contribute to the development of critical thinking, problem-solving and facilitate conclusion-reaching. With respect to the importance of comparison in geography education, the aim of the research in this paper was set: to determine the attitudes of elementary school students towards comparison as a method of teaching. The attitudes of the students towards comparison in teaching geography and the acquisition of geographical knowledge, as well as in the increase of the students’ interest, motivation and activation in the process were examined. It was also investigated whether there is a difference in students’ attitudes towards the method of comparison in relation to gender, Geography grade and the grade they attend. The surveying method was used. The results of the research show that the students expressed mostly positive attitudes towards the use of the comparison method in teaching geography. The results are also indicative of the importance of a geographical map in the application of the comparison method.

Aromatic plants and essential oils have many applications in medicine, pharmaceuticals, cosmetics, and the food industry. The essential oil of the flowering aerial parts of Achillea grandifolia, obtained by hydrodistillation, was analyzed for its constituents and investigated for antimicrobial and radical scavenging activity. The essential oil was characterized by a high amount of oxygenated monoterpenes (72.7%) with 1,8-cineole (29.2%) and camphor (23.4%) being the most abundant. Sesquiterpenes were present in smaller quantities (4.8%). Antimicrobial activity was tested against eight ATCC bacterial strains and two ATCC strains of Candida albicans. The essential oil exhibited highly pronounced antimicrobial activity against Micrococcus luteus with a MIC value of 3.50 μg/mL, as well as significant antimicrobial activity (

Cinnamon bark is used worldwide due to its characteristic flavour and medicinal properties. Ceylon cinnamon or “true” cinnamon bark refers to the dried inner bark of the shoots of Cinnamomum verum J. Presl, originated from Sri Lanka. The bark of some other species of this genus, Cinnamomum cassia Blume (Chinese cinnamon), C. burmanni (Nees & T. Nees) Blume (Indonesian cinnamon), and C. loureiroi Nees (Saigon cinnamon) are also marketed and sold as cinnamon. They are characterised by a significantly higher amount of coumarin compared to Ceylon cinnamon bark. Since coumarin may be potentially hepatotoxic, the aim of this study was to determine coumarin level in commercial samples of cinnamon bark and in cinnamon-containing dietary supplements present on the Serbian market. HPLC analysis showed lowest coumarin content in Ceylon cinnamon bark samples (0.08-0.15 mg g-1), whereas other samples contained a significantly higher amounts of coumarin (1.38-5.80 mg g-1). Cinnamon based dietary supplements contained 0.007-1.19 mg coumarin/tablet. The obtained results indicate that the majority of commercial samples of cinnamon bark on the Serbian market do not originate from the Ceylon cinnamon but from other species of this genus, and that consumed amount of certain products should be taken into account since the tolerable daily intake of coumarin is limited.

Juglans nigra (Black walnut) is a source of health-promoting biologically active compounds used in traditional medicine. The investigation of bioactive compounds in black walnut could lead to its broader application, as well as application of its by-products. Therefore, this study aimed to characterize J. nigra nut and green husk based on chemical analysis of their petroleum ether and ethanol extracts obtained by ultrasonic and reflux extraction methods, respectively. Different extract fractions were tested for their antimicrobial activities using Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa), Gram-positive bacteria (Enterococcus faecalis Staphylococcus aureus) and yeast (reference strain and clinical isolates of Candida albicans). Ethanol extracts analysis, performed by High Performance Liquid Chromatography (HPLC), singled out the ellagic acid as the most dominant compound in nut (55.0 ? 1.3 ? 10-3 kg m-3) and green husk (114.1 ? 0.5? 10-3 kg m-3) extracts. Non-polar compounds were evaluated using Gas Chromatography (GC) analysis of petroleum ether extracts. Juglans nigra nut and green husk contained two saturated fatty acids, palmitic acid (C16:0) and stearic acid (C18:0), then, monounsaturated fatty acids, palmitoleic (C16:1n-7), oleic (C18:1n-9), and vaccenic acid (C18:1n-7), as well as polyunsaturated fatty acids, linoleic (C18:2n-6), ?-linolenic (C18:3n-6), and ?-linolenic (C18:3n-3) acids. Ethanol extracts of both J. nigra nut and green husk showed antimicrobial activity against C. albicans which is the most common cause of yeast infections.

Posljednjih nekoliko decenija, zahvaljujući istraživanjima implicitnih teorija o kreativnosti, dobili smo uvid u lična shvatanja, kako laika tako i eksperata, o kreativnosti. U ovom radu bavili smo se ne samo uvjerenjima ispitanika već i time da li su, i ako jesu, u kojoj mjeri su kod nastavnika izražena pogrešna uvjerenja o kreativnosti. U ispitivanju je učestvovalo ukupno 106 ispitanika (79,25% žene; 20,75% muškarci). Uzorak su činili profesori, nastavnici i učitelji osnovnih i srednjih škola u Nikšiću. Podaci su prikupljeni upitnikom o sociodemografskim varijablama i anketom napravljenom za svrhu ovog istraživanja, koja se sastojala od 16 tvrdnji o najčešćim pogrešnim uvjerenjima o kreativnosti, gdje se stepen slaganja mogao izraziti zaokruživanjem jedne vrijednosti na petostepenoj skali Likertovog tipa. U obradi podataka korišteni su t-testovi za jedan uzorak. Rezultati istraživanja ukazuju na to da su kod profesora, nastavnika i učitelja najizraženija sljedeća pogrešna uvjerenja o kreativnosti: djeca su kreativnija od odraslih (t = 6,884; p ,001; d = 0,669), mnogi kreativni produkti ostaju nepriznati u vrijeme njihovog stvaranja i bivaju otkriveni i priznati tek godinama kasnije (t = 8,743; p ,001; d = 1,076), kreativnost predstavlja jedinstven izraz unutrašnje duše pojedinca (t = 11,078; p ,001; d = 1,076) i kreativnost je spontana inspiracija koja nije planirana ili organizovana, tj. dešava se spontano (t = 5,336; p ,001; d = 0,518). Dobijeni rezultati ukazuju na opravdanost nastavka studije sa ciljem konstrukcije skale za ispitivanje izraženosti uvjerenja o kreativnosti.

Ovim istraživanjem se nastoji, na kvalitativan način, doći do saznanja kako nastavnici opažaju darovite učenike. Interesuje nas koje su to intelektualne, motivacione, ličnosne i emocionalne, a potom i socijalne karakteristike darovitih učenika prema viđenju nastavnika. Istraživanje je obuhvatilo ukupno 15 nastavnika, iz dvije osnovne škole sa područja grada Doboja, u Bosni i Hercegovini. Za dobijanje kvalitativnih podataka korišten je polustrukturisani intervju. Rezultati dobijeni tematskom analizom ukazuju da nastavnici darovite učenike percipiraju kao učenike sa visokim intelektualnim sposobnostima, visokom unutrašnjom motivacijom, širokim interesovanjima i određenim nivoima aspiracije. Samouvjereni su, arogantni, skloni perfekcionizmu i autonomiji. Odlikuje ih ekstroverzija, ali i introverzija. Mogu biti emocionalno hladni, osjetljivi, vulnerabilni, ali i stabilni i impulsivni. U pogledu socijalnih karakteristika su ili društveni ili usamljeni. Dobijeni rezultati upućuju da nastavnici dobro prepoznaju navedene karakteristike, ali u kontekstu škole i školskih postignuća nastavnici darovitim učenicima smatraju učenike koji su dobro uklopljeni u školski sistem. Ovakvi nalazi pružaju mogućnost budućim istraživanjima da percepciju nastavnika detaljnije ispitaju i ustanove potencijalne razloge za takvu sliku darovitih učenika.

In the present research, the chemical composition of aerial parts of cultivated citral chemotype of Thymus pannonicus All. (Lamiaceae), growing at different soil conditions was studied. Two plant lineages were unfertilized or treated using either nitrogen or phosphorus fertilizers at different concentration levels. Essential oil (EO) content, EO composition, and composition of hydromethanol extracts (MEs) were compared to the results obtained for wild growing plants in order to evaluate the potential of T. pannonicus for its industrial production. EO content (%, V/m) in cultivated samples varied between 0.32-0.75% and 0.62–1.05% within two examined lineages, whereas the herbs from the natural habitat contained 0.49–1.29% of EO. GC-FID/MS analysis showed that citral, i.e. mixture of isomers geranial and neral, was the most abundant constituent in all EOs (51.90–81.96%). HPLC analysis revealed that total caffeic acid derivatives, with rosmarinic acid (RA) as the most abundant compound in all extracts, as well as total apigenin glycosides contents were significantly higher in the extracts originating from the cultivated herbs (112.15–184.94 mg/g and 5.08–15.29 mg/g, respectively), in comparison to the extracts of the herbs from the natural habitats (67.61–98.75 mg/g and 1.17–1.32 mg/g). In general, the composition of cultivated herbs varied less than that of samples originating from natural habitats, indicating that controlled cultivation of citral chemotype of T. pannonicus can provide herbal drug with favourable characteristics. Only minor differences were observed between cultivated samples treated with different fertilizers.

The extraction yield of Juglans nigra L. leaves was assessed at different ethanol concentrations (0–96% (v/v)) and solvent-to-solid ratios (5–20 kg kg−1). The response surface methodology (RSM) and artificial neural network with genetic algorithms (ANN-GA) were developed to optimize the extraction variables. The RSM and ANN-GA models determined 50% (v/v) ethanol concentration and 20 kg kg−1 solvent-to-solid ratio as optimal conditions, ensuring an extraction yield of 27.69 and 27.19 g 100 g−1 of dry leaves. The phenolic compounds in optimal extract were quantified: 3-O-caffeoylquinic acid (2.27 mg g−1of dry leaves), quercetin-3-O-galactoside (10.99 mg g−1 of dry leaves) and quercetin-3-O-rhamnoside (15.07 mg g−1of dry leaves) using high-performance liquid chromatography (HPLC). The minerals in optimal extract were quantified: macro-elements (the relative order by content was: K > Mg > Ca) using inductively coupled plasma optical emission spectrometry (ICP-OES) and micro-elements (the relative order by content was: Zn > Rb > Mn > I>Sr > Ni > Cu > Co > V > Ag > Se) using inductively coupled plasma mass spectrometry (ICP-MS). The extraction coefficients for minerals were determined and were highest for K (64.3%) and I (53.5%). Optimization of extraction process resulted in high extraction yield from J. nigra leaves and optimal extract containing different phytochemical compounds. This is the peer-reviewed version of the article: Rajković, K. M.; Vasić, M.; Drobac, M.; Mutić, J.; Jeremić, S.; Simić, V.; Stanković, J. Optimization of Extraction Yield and Chemical Characterization of Optimal Extract from Juglans Nigra L. Leaves. Chemical Engineering Research and Design 2020, 157, 25–33. [https://doi.org/10.1016/j.cherd.2020.03.002]

The composition of leaf and flower essential oils of eight Heracleum taxa (populations collected in Serbia, Montenegro, North Macedonia and Slovenia) was statistically analyzed to evaluate its chemosystematic significance. Investigated taxa included H. orphanidis and the representatives of H. sphondylium group: H. sphondylium, H. sibiricum, H. montanum, H. ternatum, H. pyrenaicum subsp. pollinianum, H. pyrenaicum subsp. orsinii and H. verticillatum. Hydrodistilled essential oils were analyzed by GC–FID and GC–MS. Chemosystematic significance was evaluated using multivariate statistics: principal component analysis (PCA), non-metric multidimensional scaling (NMDS) and unweighted pair-group arithmetic averages clustering (UPGMA). Statistical analyses included our previously published data on the composition of essential oils of eight leaf and three flower samples, as well as the data on the composition of the oils of additional eight leaf and five flower samples obtained in the current work. Leaf and flower essential oils of H. sphondylium group members were dominated by various sesquiterpenes, phenylpropanoids and/or monoterpenes. Heracleum orphanidis leaf and flower essential oils were rich in aliphatic esters, mostly octyl acetate. Statistical analysis of the composition of leaf essential oils, as well as of flower oils, demonstrated the grouping of investigated populations of these Heracleum taxa according to their systematics, i.e., separation of H. orphanidis from the representatives of H. sphondylium group, and grouping of H. pyrenaicum subspecies within this group. Morphologically related H. sibiricum and H. ternatum were closely located in PCA and NMDS and in UPGMA even shared the same cluster.

The plants of the genus Armoracia P. Gaertn., B. Mey. & Scherb. (Brassicaceae) contain glucosinolates, which volatile hydrolysis products are known for their antimicrobial activity. In this work, the composition and antimicrobial activity of the root volatiles of wild-growing Armoracia macrocarpa (Waldst. & Kit.) Kit. ex Baumg., obtained by hydrodistillation, were analysed for the first time. These results were compared with those of widely used horseradish, A. rusticana P. Gaertn., B. Mey. & Scherb. Additionally, the possibility of the propagation of A. macrocarpa from the roots, and impact of its cultivation on the composition and antimicrobial activity of the root volatiles were investigated. The GC-FID and GC–MS analysis revealed that all A. macrocarpa root volatile fractions were dominated by berteroin (55.0–59.0%) and lesquerellin (34.1–36.4%), and were significantly different from horseradish root volatile fraction. In microdilution method, A. macrocarpa volatile fractions exhibited weak/no antibacterial activity, while their effect against standard strain (MICs = 4.8–9.3 μg/mL) and clinical isolates (MICs = 25–119 μg/mL) of Candida albicans was significant. Horseradish volatiles exhibited better antibacterial and slightly weaker anticandidal activity. Armoracia macrocarpa represents a new source of raw materials with potential use in pharmaceutical and food industries, as well as in cookery. Preliminary results indicated the possibility of its cultivation, which is required for its preservation and sustainable usage.

In a search for novel food additives/functional food ingredients, chemical composition, and antioxidant and antimicrobial activities of Thymus dacicus Borbas (Lamiaceae) flowering aerial parts dry methanol extract and essential oil were tested for the first time. HPLC analysis of the extract revealed that among dominant polyphenols, rosmarinic acid was the most abundant (71.33 ± 1.11 mg/g). Monoterpenes geranial (34.4%) and neral (23.9%) prevailed in the oil, analyzed by GC and GC–MS. The extract showed pronounced antioxidant activity, with FRAP value of 5.16 ± 0.19 mmol Fe 2+ /g, OH radical SC 50 of 18.85 μg/mL (determined by 2-deoxyribose assay) and DPPH radical SC 50 of 28.04 μg/mL. Eight out of ten tested microbial strains were susceptible to the oil in microdilution test. Furthermore, for the combination of the oil and vancomycin/gentamicin, synergism was demonstrated against MRSA, Klebsiella pneumoniae and Escherichia coli in checkerboard test. Obtained results encourage further research of this herb as potentially beneficial ingredient in foodstuffs.

Geranium species are widely used in traditional medicine of Balkan. The aim of this work was to investigate and compare chemical composition of volatile fractions obtained by hydrodistillation from aerial parts of G. macrorrhizum, G. phaeum, G. sanguineum, G. robertianum, G. palustre, G. pyrenaicum, G. columbinum and G. lucidum as well as from underground parts of G. macrorrhizum and G. phaeum, originated from Vlasina plateau in South Eastern Serbia. The volatiles were analyzed using GC/MS and GC-FID. G. palustre volatiles have been studied for the first time with β-selinene (18.6 %) as a characteristic compound. The cluster analysis revealed separation of volatiles into two main groups. Volatile fractions of G. macrorrhizum were separated from all other samples due to high sesquiterpene content (92.3 % in aerial and 94.6 % in underground parts). The volatile fractions of other samples were mainly composed of sesquiterpenes (10.8–61.8 %), diterpenes (12.9–43.0 %) and fatty acids and their derivatives (6.6–21.6 %) with the exception of volatile fraction of G. phaeum underground parts which was dominated only by fatty acids and their derivatives (76.6 %). The results presented in this article contribute to the knowledge on the chemistry of this genus and advances the knowledge on flora of southeast Serbia.

For essential oils of roots, leaves, flowers and fruits of eight Heracleum taxa (H.sphondylium, H. sibiricum, H. montanum, H. ternatum, H. pyrenaicum subsp. pollinianum, H. pyrenaicum subsp. orsinii, H. verticillatum and H. orphanidis), we previously demonstrated antimicrobial, cytotoxic (selective to cancer cells) and/or antioxidant activities. In this work, for these essential oils maximum daily intake related to total furanocoumarins (FCs) content was estimated, according to Committee on Herbal Medicinal Products of European Medicines Agency (EMA/HMPC) recommendations. FCs were quantified using gas chromatography, and their sum equivalent to xanthotoxin (8-methoxypsoralen, 8-MOP) was calculated. It was shown that daily intake, not contributing significantly to overall risk (equivalent to intake of 1.5 mg FCs) for root essential oils was in the range of 1.94-5.23 mL, for fruit oils of 5.23-15.68 mL and for leaf or flower oils of 2.90-15.68 mL. Daily intake, not posing any unacceptable risk (equivalent to intake of 15 μg FCs) for root oils was in the range of 0.02-0.05 mL, for fruit oils of 0.05-0.16 mL and for leaf or flower oils of 0.03-0.16 mL. This work demonstrates the application of current EMA/HMPC recommendations, in order to establish safety profile of herbal preparations containing FCs. Za etarska ulja korena, listova, cvasti i plodova osam taksona roda Heracleum (H. sphondylium, H. sibiricum, H. montanum, H. ternatum, H. pyrenaicum subsp. pollinianum, H. pyrenaicum subsp. orsinii, H. verticillatum i H. orphanidis) prethodno su pokazane antimikrobna, citotoksična (selektivna prema tumorskim ćelijama) i/ili antioksidantna aktivnost. Cilj ovog rada bio je da se ustanovi maksimalni dozvoljeni dnevni unos etarskih ulja u odnosu na utvrđeni sadržaj ukupnih furanokumarina, u skladu sa preporukama Komiteta za biljne lekovite proizvode Evropske agencije za lekove (EMA/HMPC). Furanokumarini su kvantifikovani gasnom hromatografijom i njihov ukupni sadržaj izražen je kao ksantotoksin (8- metoksipsoralen, 8-MOP). Dnevni unos ispitivanih etarskih ulja koji ne doprinosi značajno ukupnom riziku (ekvivalentan unosu 1,5 mg furanokumarina izraženih kao 8-MOP), kreće se u opsegu 1,94-5,23 mL za etarska ulja korena, 5,23-15,68 mL za ulja plodova i 2,90-15,68 mL za ulja listova ili cvasti. Dnevni unos etarskih ulja koji ne predstavlja nikakav rizik (ekvivalentan unosu 15 μg furanokumarina izraženih kao 8-MOP), kreće se u opsegu 0,02-0,05 mL za etarska ulja korena, 0,05-0,16 mL za ulja plodova i 0,03-0,16 mL za ulja listova ili cvasti. Ovaj rad demonstrira primenu aktuelnih EMA/HMPC preporuka u cilju utvrđivanja bezbednosnog profila biljnih preparata u kojima su detektovani furanokumarini.

The composition of essential oils obtained by hydrodistillation from dried stems and inflorescences of Cymbopogon nervatus (Hochst.) Chiov. (Poaceae) growing wild in Western and Eastern Sudan and collected at two different periods, was analyzed by GC and GC–MS. The most dominant constituents in the oils were p -menthadienols, trans - p -mentha-1(7),8-dien-2-ol (19.9–32.6%), cis-p -mentha-1(7),8-dien-2-ol (18.9–23.3%), trans-p -mentha-2,8-dien-1-ol (10.6–21.0%) and cis-p -mentha-2,8-dien-1-ol (8.1–10.4%). The essential oil of inflorescence from the Western Sudan was tested for spasmolytic activity using three different experimental models: against spontaneous contractions, and contractions induced with ACh and KCl. The oil exhibited strong, significant and concentration-dependent spasmolytic activity. Applied in concentration of 200 μg/ml, the oil showed 88.44 ± 16.78% of maximal spasmolytic effect of atropine (6.4 μM) against spontaneous contraction. Essential oil (90 μg/ml) inhibited ACh-induced contractions as well, and reduced the effect of the highest applied concentration of ACh to 37.29 ± 16.16%. Additionally, the oil exhibited strong activity against contractions induced with KCl (80 mM) and in concentration of 200 μg/ml, completely abolished the contractile effect of KCl. The essential oil of inflorescence from the Western Sudan was also tested for antioxidant activity by DPPH assay and showed moderate anti-DPPH activity (SC 50 23.8 μl/ml).

The compositions of hydrodistillated essential oils of Laser trilobum (L.) Borkh. rhizomes and fruits from Serbia, were investigated using GC and GC/MS. In the dark-blue rhizome oil forty-six compounds (93.1% of the total oil) were identified, with α-pinene (31.5%), γ-terpinene (9.0%), p-cymene (7.9%), β-pinene (6.1%) and 1,4-dimethylazulene (6.0%) as the major components. In the colorless fruits oil, twenty components (96.8% of the total oil) were identified, and the main constituents were limonene (51.6%) and perillaldehyde (26.8%). The antimicrobial activity of the oils was tested using the broth microdilution method against nine bacterial and two fungal strains. The oils revealed significant antimicrobial effect, mainly better than that of thymol, used as a reference compound. The strongest activity was recorded for the rhizome oil against Escherichia coli, Klebsiella pneumoniae and Candida albicans (MICs=25 μg/mL), and the fruit oil against C. albicans ATCC 10259 (MIC=12.5 μg/mL).

The composition of the essential oils and the furanocoumarin profiles of CH2 Cl2 extracts from underground parts and fruits of nine Heracleum taxa (Apiaceae) from Southeastern Europe were statistically analyzed to evaluate their chemosystematic significance. Eight investigated taxa (H. orphanidis and members of H. sphondylium group: H. sphondylium, H. sibiricum, H. montanum, H. ternatum, H. pyrenaicum subsp. pollinianum, H. pyrenaicum subsp. orsinii and H. verticillatum) belong to the type section of the genus. Additionally analyzed taxon, H. austriacum subsp. siifolium, belongs to H. sect. Wendia. Qualitative and quantitative analysis of the essential oils was performed by GC-FID and GC/MS, and of the furanocoumarins by LC/MS. Furanocoumarins were identified using standards and/or based on UV, MS, 1 H- and ROESY NMR spectra. Multivariate statistics (PCA, nMDS, UPGMA) of analyzed metabolites showed that the investigated taxa were grouped according to their taxonomic classification. PCA revealed the significance of some monoterpenes and (Z)-falcarinol of the underground parts oils, octyl esters of the fruit oils, and most of the total of 12 identified furanocoumarins.

Many Heracleum L. taxa (Apiaceae) are used as food and spices, and in traditional medicine. In this work, the chemical composition of Heracleum pyrenaicum subsp. orsinii (Guss.) F. Pedrotti and Pignatti root, leaf and fruit essential oils, their antimicrobial activity and cytotoxic effect on malignant and normal cells were investigated for the first time. The composition of the oils was analyzed by GC and GC–MS. Monoterpenes prevailed in the root oil, with β-pinene (38.6%) being dominant, while in the leaf oil, sesquiterpenes, mostly (E)-nerolidol (20.5%) and (E)-caryophyllene (17.0%), were the most abundant constituents. The fruit oil contained the majority of aliphatic esters, mainly octyl acetate (36.8%) and octyl hexanoate (22.1%). The antimicrobial activity was determined by microdilution method against eight bacteria and eight fungi (standard strains, clinical or food isolates). The best antibacterial activity, better than the activity of ampicillin, was shown by the root oil against Salmonella typhimurium, Escherichia coli and Pseudomonas aeruginosa. The strongest antifungal activity, stronger than the activity of ketoconazole, was exhibited by the leaf and root oils against Trichoderma viride, and by the root oil against Aspergillus ochraceus. The cytotoxic effect of the oils, determined by MTT test, was prominent against malignant HeLa, LS174 and A549 cells (IC50=6.49–14.56μg/mL). On the other hand, the oils did not show toxicity against normal MRC-5 cells at tested concentrations (IC50>200.00μg/mL). It can be concluded that investigated H. pyrenaicum subsp. orsinii oils represent potential new raw materials for food and pharmaceutical industry.

Herbal drugs are often administered in the form of monocomponent teas or herbal tea mixtures (tea blends). In order to assess the quality of herbal drugs that are components of the monocomponent teas commercially available on marketplaces in Belgrade, we analyzed 4 samples of Melissa leaf (Melissae folium), 6 samples of Wild thyme, (Serpylli herba), 6 samples of Yarrow (Millefolii herba), 4 samples of Wormwood (Absinthii herba) and 6 samples of Juniper (Juniperi pseudo-fructus), according to the procedures given in European Pharmacopoea. Macroscopic and microscopic features of investigated herbal drugs, as well as their TLC profiles, were in agreement with corresponding data described in Ph. Eur. One sample of Melissa leaf, one sample of Wild thyme and two samples of Juniper, did not fullfil the requirements of general tests. Due to inappropriate size of cut, test on foreign matter could not be performed for some samples. Concerning specific quality, all samples of Melissa leaf, Wild thyme, Wormwood and Juniper were in accordance with Ph. Eur. Regarding the content of essential oil, Yarrow samples met the requirements of Ph. Eur., but the proazulenes content in five samples was lower than required. In conlcusion, the results of these analyses point to high medicinal potential of the most herbal drugs studied. On the other hand, unsatisfying general quality and inappropriate size of cut of several samples could be related to inadequate processing and handling of plant material. Biljne droge se vrlo često koriste u obliku monokomponentnih i višekomponentnih čajeva. U cilju ispitivanja kvaliteta biljnih droga koje se mogu naći na pijacama Beograda u obliku monokomponentnih čajeva, analizirana su po četiri uzorka lista matičnjaka (Melissae folium) i herbe belog pelena (Absinthii herba), i po šest uzoraka herbe majčine dušice (Serpylli herba), herbe hajdučke trave (Millefolii herba) i šišarica kleke (Juniperi pseudo-fructus), prema propisu Ph. Eur. 7.0. Makroskopske i mikroskopske karakteristike ispitivanih droga, kao i rezultati TLC analize bili su u skladu sa odgovarajućim podacima datim u monografijama Farmakopeje. U pogledu opšteg kvaliteta, zahteve nisu ispunili jedan uzorak lista matičnjaka, jedan uzorak herbe majčine dušice i dva uzorka šišarica kleke. Zbog neodgovarajućeg stepena usitnjenosti pojedine uzorke nije bilo moguće ispitati u pogledu stranih primesa. Zahteve za specifični kvalitet ispunili su svi uzorci lista matičnjaka, herbe majčine dušice, herbe belog pelena i šišarica kleke. Uzorci herbe hajdučke trave sadržali su odgovarajuću količinu etarskog ulja, ali je sadržaj proazulena u pet uzoraka bio niži od propisanog. Rezultati ispitivanja specifičnog kvaliteta ukazuju na visok lekoviti potencijal većine ispitivanih biljnih droga. Sa druge strane, neispravnost u pogledu opšteg kvaliteta i neodgovarajući stepen usitnjenosti pojedinih uzoraka može se dovesti u vezu sa neadekvatnim rukovanjem biljnim materijalom.

During the past few years, the isophylloid Campanula pyramidalis complex has been the subject of studies aimed at an improved understanding of the relationships within it. The center of distribution of the C. pyramidalis complex is in the Balkan Peninsula with some smaller parts of the area located in the south Apennines. Although 21 taxa of the C. pyramidalis complex were described, only four species are accepted: C. pyramidalis, C. versicolor, C. secundiflora and C. austroadriatica. In the present study, we propose compounds of the methanolic leaf extract as possible chemotaxonomic markers for the C. pyramidalis complex. Eleven flavonoids and two phenolic acids were detected in leaf extract using high-performance liquid chromatography with diode-array detection analysis. The investigated taxa of the C. pyramidalis complex differ in terms of the composition of the methanolic leaf extract. clustering of investigated taxa is not completely consistent with the previously reported molecular and morphometric data.

Vascular stents are general medical devices of class III or IIb, which are placed along the walls of the constricted coronary and peripheral blood vessels thus keeping them viable. According to the mechanism of expansion, stents may be balloon-expandable or self-expanding. Depending on the geometry they can be classified into: coil stents, open-cell modular stents and multi-cell closed cell stents. The most important characteristic of vascular stents is their flexibility, but a number of additional requirements must also be met: high radial strength, low elastic deformation, small diameter, the possibility of monitoring through the bloodstream, minimum subsequent shortening, minimum elastic longitudinal deformation, and the optimal retention at target site. Materials for production of stents must be biologically inert, visible by radiological techniques, biocompatible, corrosion-resistant and resistant to stress due to blood flow. The most significant adverse event after stent implantation is the occurrence of restenosis, which is most efficiently overcome by the application of drug releasing stents. These stents are composed of three parts: stent platform, drug carrier and a drug that inhibits neointimal hyperplasia - paclitaxel and limuses (sirolimus, everolimus, zotarolimus, tacrolimus, pimecrolimus, etc.). In recent years, stents with surfaces coated with substances that accelerate endothelialisation and thus reduce thrombosis have been developed. The latest approach is represented by a stent with the lumen coated with CD34 antibody, and the outer side coated with sirolimus. By using these devices a double effect is achieved: acceleration of endothelialisation and inhibition of neointimal hyperplasia. Vaskularni stentovi su opšta medicinska sredstva klase III ili IIb, koja se postavljaju uz zidove koronarnih i perifernih krvnih sudova kada postoji suženje i na taj način ih održavaju prohodnim. Prema mehanizmu ekspanzije, stentovi mogu biti balon-ekspandirajući ili samoekspandirajući, a u zavisnosti od geometrijskog oblika dele se na: stentove u obliku spirale, modularne stentove sa otvorenim ćelijama i multićelijske stentove sa zatvorenim ćelijama. Najznačajnija karakteristika vaskularnih stentova je fleksibilnost, ali moraju da ispunjavaju i čitav niz dodatnih zahteva, kao što su: velika radijalna snaga, nizak stepen elastične deformacije, mali prečnik, mogućnost praćenja kroz krvotok, minimalno naknadno skraćivanje, minimalna elastična longitudinalna deformacija i optimalno zadržavanje na ciljnom mestu. Materijali za proizvodnju stentova moraju biti biološki inertni, vidljivi radiološkim tehnikama, biokompatibilni, otporni na koroziju i otporni na stres zbog protoka krvi. Najznačajniji neželjeni događaj nakon ugradnje stentova je pojava restenoze, koja se najefikasnije prevazilazi primenom stentova koji oslobađaju lek. Ovi stentovi se sastoje iz tri dela: platforme stenta, nosača leka i leka koji inhibira neointimalnu hiperplaziju, paklitaksel i limusi (sirolimus, everolimus, zotarolimus, takrolimus, pimekrolimus i dr). Poslednjih godina razvijeni su i stentovi kod kojih se površine oblažu supstancama koje ubrzavaju endotelizaciju i tako smanjuju trombozu. Najnoviji pristup predstavlja stent kod koga se lumen stenta oblaže CD34 antitelom, a spoljašnja strana sirolimusom. Primenom ovih stentova postiže se dvostruki efekat: ubrzavanje endotelizacije i inhibicija neointimalne hiperplazije.

In the paper, we have analysed curricula of the several subjects in lower grades of the primary school in Republika Srpska. Blum's taxonomy was used in the analysis (cognitive and psycho-motor area) and SMART system of evaluating learning outcomes. There have been three subject areas included (the world around us, speech, expression, crating and rhythm, sport and music), in the first grade ten subjects (Serbian, Mathematics, Natural and Social Sciences, etc.), and pedagogical work with the class, which are being studied from the second to the fifth class. The aim of the analysis was to determine knowledge and skills which students should gain in the lower grades of the primary school. We have achieved this aim by determination of the outcomes adequacy based on the above stated criteria, levels of knowledge and skills, which are required, as well as estimation of how much each teaching unit requires certain level of students' knowledge and whether there is the trend of more requirements when passing one grade and entering the other. The results of the analysis show that students are required the lowest level of understanding and gaining (on average 60% of outcomes are on this level), and higher levels require only application (about 15%). This relation varies, depending on the subject. When we talk about skills, they are at the level of precision, and this is sufficient for this age. The rising requirements for students, through learning outcomes have not been observed, because from grade to grade the outcomes are on the same level. This means that we do not work sufficiently at the quality improvement, but on quantity improvement of knowledge.

Fatty acids (FAs), sterols, and triterpenes of dichloromethane extracts of the fruits of 8 Heracleum L. taxa (Apiaceae) from southeastern Europe were investigated by gas chromatography with flame ionization detector and gas chromatography with mass spectrometry. In order to analyze the FAs, their volatile methyl esters were obtained by saponification and subsequent esterification of oily supernatants of the fruit extracts. Dominant was petroselinic acid (42.8%-56.5%), followed by linoleic (20.3%-33.3%) and oleic acids (12.3%-13.7%). Sterols and triterpenes were analyzed as volatile derivatives obtained by the silanization of residual unsaponifiable fractions. Among them, the most abundant was β-sitosterol (44.9%-56.9%), followed by stigmasterol (15.7%-25.0%), Δ7-stigmastenol (6.6%-12.5%), and campesterol (5.2%-8.1%). The quantity of petroselinic acid was also determined by the external standard method (298.8-433.4 mg/g of oily supernatant). The obtained results show that the investigated plants are potential valuable sources of the compounds utilized in different industries.

Many Heracleum L. taxa (Apiaceae) are used as food and spices, and in traditional medicine. In this work, the chemical composition of Heracleum pyrenaicum subsp. orsinii (Guss.) F. Pedrotti and Pignatti root, leaf and fruit essential oils, their antimicrobial activity and cytotoxic effect on malignant and normal cells were investigated for the first time. The composition of the oils was analyzed by GC and GC–MS. Monoterpenes prevailed in the root oil, with β-pinene (38.6%) being dominant, while in the leaf oil, sesquiterpenes, mostly (E)-nerolidol (20.5%) and (E)-caryophyllene (17.0%), were the most abundant constituents. The fruit oil contained the majority of aliphatic esters, mainly octyl acetate (36.8%) and octyl hexanoate (22.1%). The antimicrobial activity was determined by microdilution method against eight bacteria and eight fungi (standard strains, clinical or food isolates). The best antibacterial activity, better than the activity of ampicillin, was shown by the root oil against Salmonella typhimurium, Escherichia coli and Pseudomonas aeruginosa. The strongest antifungal activity, stronger than the activity of ketoconazole, was exhibited by the leaf and root oils against Trichoderma viride, and by the root oil against Aspergillus ochraceus. The cytotoxic effect of the oils, determined by MTT test, was prominent against malignant HeLa, LS174 and A549 cells (IC50 = 6.49–14.56 μg/mL). On the other hand, the oils did not show toxicity against normal MRC-5 cells at tested concentrations (IC50 > 200.00 μg/mL). It can be concluded that investigated H. pyrenaicum subsp. orsinii oils represent potential new raw materials for food and pharmaceutical industry.

Although some widespread, native cow parsnips (Heracleum L. spp., Apiaceae) had broad medicinal and culinary applications throughout history, the knowledge about their volatile constituents is insufficient. This work investigates the composition and bioactivities of H. sphondylium L. (HSPH), H. sibiricum L. (HSIB) and H. montanum Schleich. ex Gaudin (HMON) essential oils. The composition was tested by GC and GC-MS. (Z)-β-Ocimene was the most abundant in HSPH (28.9%) and HMON (20.4%) root oils, while in HSIB root oil, β-pinene (26.2%), methyl eugenol (22.3%) and elemicin (25.6%) prevailed. Leaf and flower oils were dominated by various sesquiterpenes (germacrene D, β-sesquiphellandrene, (E)-β-farnesene and/or (E)-caryophyllene) and/or phenylpropanoids (apiole, methyl eugenol, elemicin and/or (Z)-isoelemicin). Octyl acetate (57.5-67.1%) was the main constituent of all fruit oils. The antimicrobial activity was screened by a microdilution method against eight bacteria and eight fungi. The strongest antimicrobial effect, in several cases better than the activity of antibiotics, was shown by HSPH (MICs = 0.12-3.30 mg mL-1) and HMON (MICs = 0.10-1.30 mg mL-1) flower oils against bacteria, and HSIB fruit oil against fungi (MICs = 0.15-0.40 mg mL-1). The MTT test revealed that the oils were not or weakly cytotoxic against human malignant HeLa, LS174 and/or A549 cells (except HSPH root oil; IC50 = 5.72-24.31 μg mL-1) and that tested oils were not toxic against human normal MRC-5 cells (at 200.00 μg mL-1). Significant activity observed against microorganisms that are the common cause of foodborne diseases, food contamination and/or hospital-acquired infections justifies certain traditional uses of the investigated plants and represents a good basis for further research of these Heracleum oils.

The objective of this research was to analyze the chemical composition, antimicrobial and cytotoxic activity of Heracleun pyrenaicum subsp. pollinianum (Bertol.) F. PedrottiPignatti (HPP) and H. orphanidis Boiss. (HO) essential oils. The composition of the oils was analyzed by GC and GC-MS. β-Pinene (35.1%) was the most abundant compound in HPP root oil, while (Z)-falcarinol (80.0%) dominated in HO root oil. (E)-Nerolidol (28.5%) was the main constituent in HPP leaf oil. HPP fruit oil, as well as HO leaf and fruit oils mainly contained aliphatic esters, mostly octyl acetate (50.5-84.5%). Antimicrobial screening was performed by microdilution method against eight bacterial and eight fungal strains. The strongest antibacterial activity was shown by both root oils (MICs 0.02-0.60 mg/mL and MBCs 0.04-2.50 mg/mL for HPP, and MICs 0.02-1.25 mg/mL and MBCs 0.04-2.50 mg/mL for HO), while the best antifungal potential was exhibited by HPP fruit oil (MICs 0.30-0.60 mg/mL and MFCs 0.60-1.25 mg/mL) and HO leaf oil (MICs 0.15-0.63 mg/mL and MFCs 0.30-1.25 mg/mL). The tested root and fruit oils exhibited strong cytotoxic effect, which was determined by MTT test against HeLa (IC50 7.53-21.07 µg/mL) and LS174 (IC50 24.16-58.86 µg/mL) cell lines.

Hungarian thyme (Thymus pannonicus All., Lamiaceae) is an aromatic herb used as traditional remedy, a refreshing beverage and a food aromatizer. Herbal teas, i.e., infusions, of Hungarian thyme from eight localities in Serbia were analyzed regarding their polyphenolic and volatile composition, and tested for their antioxidant and antimicrobial activity. The total polyphenolics content, determined by the Folin-Ciocalteu method, ranged from 1122.25 to 1979.93 mg gallic acid/L. HPLC analysis revealed rosmarinic acid (367.42–1199.47 mg/L) and luteolin glucuronides as the main polyphenolics. The volatile fractions of the infusions, analyzed by static headspace extraction coupled with GC and GC–MS analyses, contained citral, 3-octanone, 1-octen-3-ol, linalool and 1,8-cineole as the dominant constituents. The antioxidant activity of the infusions was examined through the ferric reducing antioxidant power (FRAP) (68.09–124.58 mmol Fe2+/L) and the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical tests (SC50 1.32–2.96 μL/mL). The antimicrobial activity was tested by the broth microdilution method against standard strains of Gram(+) Staphylococcus aureus, Staphylococcus epidermidis, Bacillus subtilis and Enterococcus faecalis and Gram(−) bacteria Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa, and one strain of yeast Candida albicans. The infusions inhibited microbial growth in the tested concentration range (31.25–500.00 μL/mL) and the strongest activity was exhibited against the strain of C. albicans (MIC 31.25–62.50 μL/mL). The antioxidant and antimicrobial properties of the infusions were, to some extent, in correlation with the composition and content of the polyphenolic compounds, whereas the volatiles noticeably influenced the exhibited antimicrobial activity.

Behavior bioassays were conducted in the laboratory (23 ± 1°C and 50 ± 5% r.h.) using the olfactometer to determine the effects of essential oils of three plant species in the Cymbopogon genus (Lemongrass), Cymbopogon nervatus, C. proximus and C. schoenanthus, on adults of Tribolium castaneum. The effect of essential oils was compared to a commercial biopesticide based on azadirachtin at three concentrations (0.0001, 0.001 and 0.01%). The results showed that all essential oils and the azadirachtin-based biopesticide had significant (p lt 0.05) repellent effects on T. castaneum adults at all tested concentrations, except C. proximus essential oil which showed a neutral effect at the lowest concentration. The highest concentrations of the essential oils of C. nervatus and C. proximus had significantly stronger repellent effects (p lt 0.05) than the lowest concentration. At the concentrations of 0.0001 and 0.001% all tested essential oils and azadirachtin showed a similar repellent effect without statistically significant difference, whereas the oil of C. nervatus had the highest repellent effect on adults of T. castaneum at the 0.01% concentration. Considering all tested variations, the essential oils of plants of the genus Cymbopogon showed similar or stronger repellent effects on T. castaneum adults than the biopesticide based on azadirachtin. U laboratorijskim uslovima (24±1°C i 50±5% r.v.v.) je pomoću olfaktometra ispitivan uticaj etarskih ulja iz tri biljne vrste roda limunovih trava (Cymbopogon nervatus, Cymbopogon proximus i Cymbopogon schoenanthus) na ponašanje (atraktivnost/repelentnost) adulta T. castaneum. Uticaj etarskih ulja na ponašanje kestenjastog brašnara, poređen je sa uticajem biopesticida na bazi azadirahtina u tri koncentracije: 0.0001, 0.001 i 0.01%. Rezultati testiranja pokazali su da sva etarska ulja i biopesticid na bazi azadirahtina u svim ispitivanim koncentracijama ispoljavaju statistički značajan odbijajući uticaj - repelentni efekat na adulte kestenjastog brašnara, osim najniže koncentracije ulja iz biljke C. proximus koja je ispoljila neutralan efekat na brašnara. Najviša koncentracija ulja iz biljaka C. nervatus i C. proximus je ispoljila statistički značajno jači repelentni efekat u odnosu na najnižu koncentraciju. Sva ispitivana etarska ulja i azadirahtin su u koncentracijama 0.0001 i 0.001% ispoljila sličan repelentni efekat bez statistički značajne razlike, dok je ulje iz biljke C. nervatus u koncentraciji 0.01% prouzrokovalo statistički značajno jači repelentni efekat na adulte kestenjastog brašnara u odnosu na ostale ispitivane supstance. Na osnovu rezultata istraživanja sa adultima kestenjastog brašnara može se zaključiti da su etarska ulja iz tri vrste biljaka iz roda Cymbopogon ispoljila sličan i/ili jači repelentni efekat od biopesticida na bazi azadirahtina. Takođe, dobijeni rezultati pokazuju značajan potencijal upotrebe ulja ovih biljaka kao prirodnog sredstva za zaštitu uskladištenih biljnih proizvoda.

In this work, the chemical composition, antimicrobial and cytotoxic activity of Heracleum verticillatum Pančić and H. ternatum Velen. root, leaf, and fruit essential oils were investigated. The composition was analyzed by GC and GC/MS. Heracleum verticillatum and H. ternatum root oils were dominated by monoterpenes, mostly β-pinene (23.5% and 47.3%, respectively). Heracleum verticillatum leaf oil was characterized by monoterpenes, mainly limonene (20.3%), and sesquiterpenes, mostly (E)-caryophyllene (19.1%), while H. ternatum leaf oil by the high percentage of phenylpropanoids, with (Z)-isoelemicin (35.1%) being dominant constituent. Both fruit oils contained the majority of aliphatic esters, mostly octyl acetate (42.3% in H. verticillatum oil and 49.0% in H. ternatum oil). The antimicrobial activity of the oils was determined by microdilution method against eight bacterial and eight fungal strains. The strongest effect was exhibited by H. verticillatum root oil, particularly against Staphylococcus aureus, Salmonella typhimurium (MICs = 0.14 mg/ml, MBCs = 0.28 mg/ml), and Trichoderma viride (MIC = 0.05 mg/ml, MFC = 0.11 mg/ml). Cytotoxic effect was determined by MTT test against malignant HeLa, LS174, and A549 cells (IC50 = 5.9 - 146.0 μg/ml), and against normal MRC-5 cells (IC50 > 120.1 μg/ml). The best effect was exhibited by H. verticillatum root oil on A549 cells (IC50 = 5.9 μg/ml), and H. ternatum root oil against LS174 cells (IC50 = 6.7 μg/ml).

The chemical composition and antimicrobial activity of essential oils of Laserpitium latifolium and L. ochridanum were investigated. The essential oils were isolated by steam distillation and characterized by GC-FID and GC/MS analyses. All essential oils were distinguished by high contents of monoterpenes, and α-pinene was the most abundant compound in the essential oils of L. latifolium underground parts and fruits (contents of 44.4 and 44.0%, resp.). The fruit essential oil was also rich in sabinene (26.8%). Regarding the L. ochridanum essential oils, the main constituents were limonene in the fruit oil (57.7%) and sabinene in the herb oil (25.9%). The antimicrobial activity of these essential oils as well as that of L. ochridanum underground parts, whose composition was reported previously, was tested by the broth-microdilution method against four Gram-positive and three Gram-negative bacteria and two Candida albicans strains. Except the L. latifolium underground-parts essential oil, the other investigated oils showed a high antimicrobial potential against Staphylococcus aureus, S. epidermidis, Micrococcus luteus, or Candida albicans (minimal inhibitory concentrations of 13.0-73.0 μg/ml), comparable to or even higher than that of thymol, which was used as reference compound.

In this paper antinociceptive and anti-edematous effects are examined of the essential oils of the underground parts of two Balkan endemic Laserpitium species (Apiaceae), L. zernyi and L. ochridanum. Furthermore, the essential oil of the underground parts of L. ochridanum is chemically characterised by GC and GC-MS. Antinociceptive and anti-edematous effects were measured in a rat model of localized inflammation, induced by carrageenan, using apparatus for the modified paw-pressure test, and plethysmometer, respectively. The effects of both Laserpitium essential oils were measured after oral gavage administration to male Wistar rats in doses of 25, 50 and 100 mg/kg. The main constituents of L. ochridanum essential oil were: alpha-pinene (33.2%), alpha-bisabolol (10.3%) and chamazulene (14.9%). The essential oil of L. zernyi was previously shown to be rich in alpha-pinene (31.6%) and alpha-bisabolol (30.9%). Both examined essential oils produced a significant dose-dependent antinociceptive effect. The corresponding ED50 +/- SEM in producing antinociception were 45.9 +/- 4.9 mg/kg and 42.4 +/- 2.1 mg/kg for L. zernyi and L. ochridanum oil, respectively. Both essential oils also significantly reduced paw edema in a dose-dependent manner. The estimated ED50 +/- SEM values for the anti-edematous effect were 36.3 +/- 4.5 mg/kg for L. zernyi oil and 45.1 +/- 11.3 mg/kg for L. ochridanum oil. These results suggest that the essential oils of both investigated Laserpitium species may be effective against pain and edema present in various inflammatory conditions.