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© 2020, © 2020 Har Krishan Bhalla & Sons. A total of 26 experimentally obtained retention indices on a logarithmic scale (log RI) from Thymus serpyllum essential oil were used to build a robust predictive model. The selected descriptors were used as inputs of an artificial neural network model to build a predictive quantitative structure-retention time relationship model. The coefficient of determination for the training cycle was 0.977, indicating that this model could be used for the prediction of retention indices for T. serpyllum essential oil compounds. These 26 compounds comprise about 99.8 % of the total oil, but among them only 6 compounds had the average relative concentration over 1 percent: geraniol (63.4 %), nerol (or cis-geraniol) (18.9 %), geranyl acetate (4.7 %), trans- caryophyllene (4.6 %), β-bisabolene (2.0 %) and geranial (1.2 %). According to these results, it can be concluded that T. serpyllum from village Sesalac (Serbia) belongs to geraniol chemotype, in total 82.3 % in both, trans and cis forms (63.4 % and 18.9 %, respectively).

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]

Five genotypes of Turkish hazelnuts ( Corylus colurna L.) collected during the 2016 crop were analyzed for chemical composition, including total oil content, fatty acid and sterol composition. The oil yields from these kernels varied from 36.5% to 60.8%. The main fatty acids of Turkish hazelnuts kernel oils were oleic acid (79.34–83.0%) and linoleic acid (7.52–10.81%). The polyunsaturated/saturated fatty acid ratio is low, ranging from 0.87 to 1.25. Total phytosterol content ranged from 4.52 to 6.50 mg g -1 of oil. Among the eleven sterols identified and quantified, s -sitosterol was the major one with a mean percentage of 65.09%, while fucosterol and campesterol was the second and the third components of the group with mean values of 10.91 and 4.36% respectively. The antioxidant potential was evaluated by radical scavenging activity using 2.2-diphenyl-1 picrylhydrazyl (DPPH) assay.

The multicomponent mixtures consisting of herbs and fungi are commonly used for the production of alcoholic beverages with potential health-promoting effects in many Asian countries. The medicinal fungus Ganoderma lucidum is one of the most important fungi used for spirit production. Although this fungus affects the aromatic complexity of spirits, only a small number of studies have focused on investigating the influence of G. lucidum on the aromatic profile and colour of spirits. The aim of the research is to evaluate the influence of adding G. lucidum and herbal extract on final concentrations of volatile compounds and sensory quality of several distillates. In this study, distillates (grain, plum, grape and wine) were used to produce new spirit-based beverages with the fungus G. lucidum only, or with the fungus and herbal extract. Fifty-nine aroma compounds were identified by GC-MS. The aromatic profiles were strongly influenced by the primary aromas of the distillates, but the addition of G. lucidum and herbal extract enriched the volatile fraction of distillates with a range of ethyl esters, with a fruity and floral fragrance. Higher alcohols, 1-propanol, 2-isobutanol and isoamyl alcohol, were the most abundant volatile compounds in the analyzed distillates and spirits. The lightness of distillates was from 60.7 to 63.6, and with the addition of Ganoderma it significantly decreased to the range from 43.6 to 50.5. The addition of the fungus also increased the intensity of red and yellow colours. The Ganoderma spirits scored very highly in sensory evaluation (17.6–18.3), significantly better than the spirits without any additions (16.1–16.9).

Plum brandy (Slivovitz (en), Šljivovica(sr)) is an alcoholic beverage that is increasingly consumed all over the world. Its quality assessment has become of great importance. In our study, the main volatiles and aroma compounds of 108 non-aged plum brandies originating from three plum cultivars, and fermented using different conditions, were investigated. The chemical profiles obtained after two-step GC-FID-MS analysis were subjected to multivariate data analysis to reveal the peculiarity in different cultivars and fermentation process. Correlation of plum brandy chemical composition with its sensory characteristics obtained by expert commission was also performed. The utilization of PCA and OPLS-DA multivariate analysis methods on GC-FID-MS, enabled discrimination of brandy samples based on differences in plum varieties, pH of plum mash, and addition of selected yeast or enzymes during fermentation. The correlation of brandy GC-FID-MS profiles with their sensory properties was achieved by OPLS multivariate analysis. Proposed workflow confirmed the potential of GC-FID-MS in combination with multivariate data analysis that can be applied to assess the plum brandy quality.

Thirteen undescribed 24-methylene lanostane triterpenoids, named polyporenic acids E-M and fomitosides L-O, as well as seventeen known analogues, were isolated from the fruiting bodies of the mushroomFomitopsis betulina. Their structures were determined using 1D, 2D NMR, IR, and HRESIMS. Fomitoside L and fomitoside N exhibited cytotoxicity against HL60 leukemia cells (IC50=15.8 and 23.7μM, respectively).Among the known compounds, notable cytotoxicities against HL60 leukemia cells and selectivity with respect to MRC-5 healthy cells were noticed for dehydropachymic acid (IC50=10.9μM, SI 8.6), pachymic acid (IC50=11.0μM, SI 9.8), 3-epi-dehydrotumulosic acid (IC50=19.9μM, SI 5.8) and 12α-hydroxy-3α-(3′-hydroxy-4′-methoxycarbonyl-3′-methylbutyryloxy)-24-methyllanosta-8,24 (31)-dien-26-oic acid (IC50=19.2μM, SI 2.2). Graphical abstract https://ars.els-cdn.com/content/image/1-s2.0-S003194222031195X-fx1_lrg.jpg

The aim of this research was to verify the authenticity of monofloral honeys from the territory of the Republic of Serbia on the basis of physicochemical parameters routinely measured in honey quality control using multivariate analysis. Seventeen samples of monofloral honey (11 samples of acacia honey and 6 samples of sunflower honey) from the territory of the Republic of Serbia were analyzed. Physico-chemical analysis of the samples included the examination of basic quality parameters and qualitative and quantitative analyses of phenolic compounds. In the samples tested, a total of 93 phenolic compounds were tentatively identified, and 19 of them were quantified. The obtained physicochemical analysis of the data served as input for the multivariate analysis. The hеаt map, which is useful for visualizing numerical data, was used for this purpose. The obtained results showed that the applied data can serve to clearly separate acacia and sunflower honeys. Целта на ова истражување е да се потврди автентичноста на моноцветен мед од територијата на Република Србија врз основа на физичкохемиските параметри што рутински се мерат при контрола на квалитет на мед со примена на мултиваријантна анализа. Беа анализирани 17 примероци на моноцветен мед (11 примероци мед од акација и 6 примероци мед од сончоглед) од територијата на Република Србија. Физичкохемиските анализа на примероците вклучуваа испитување на основните параметри за квалитет како и квалитативна и квантитативна анализа на фенолните соединенија. Во примероците што беа тестирани, хипотетички беа идентификувани вкупно 93 фенолни соединенија, а 19 од нив беа и квантификувани. Добиените податоци од физичкохемиска анализа на овие податоци беа употребени како влезна податоци за мултиваријантна анализа. Беше применета топлинска мапа која е корисна за визуализација на нумеричките податоци. Добиените резултати покажуваат дека применетите податоци може да послужат за јасно да се раздели медот од акација и од сончоглед.

Flower strips of French Marigold are commonly used pest repellents in potato fields. However, the effect of French Marigold volatiles on potato metabolism, physiology and induced defense is unknown. Thus, a microarray transcriptome analysis was performed to study the effects of French Marigold essential oil (EO) on laboratory-grown potato. After 8 h of exposure to EO, with gas chromatography/mass spectrometry (GC/MS)-detected terpinolene and limonene as dominant compounds, 2796 transcripts were differentially expressed with fold change &gt, 2 compared to expression in controls. A slightly higher number of transcripts had suppressed expression (1493 down- vs. 1303 up-regulated). Since transcripts, annotated to different photosynthesis-related processes, were mostly down-regulated, we selected a set of 10 genes involved in the leaf starch metabolism pathway, and validated microarray patterns using quantitative reverse transcription polymerase chain reaction (RT-qPCR). Except for decreased synthesis and induced decomposition of starch granule in leaves, 8 h long EO exposure slightly elevated the accumulation of sucrose compared to glucose and fructose in subjected potato plants. An in vitro feeding bioassay with Colorado potato beetle showed that EO-induced alternations on transcriptional level and in the sugars&rsquo, metabolism caused the enhancement of feeding behavior and overall development of the tested larvae. Results of comprehensive analysis of transcriptional responses in potato exposed to French Marigold EO provide a basis for further elucidation of molecular mechanisms underlying eco-physiological interactions in companion planting cropping systems.

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.

Paper description: The headspace method (HS) enables us to study the composition of volatile compounds by using only 0.1 g of plant material and without creating artefacts during isolation. Volatiles stored in twigs are separated from needle volatiles and thus add new information to chemophenetic studies. Volatiles of 100 individuals of P. omorika collected in natural populations from seven localities were extracted for the first time using by HS. The composition was analysed statistically to assess variability and population differentiation. The dominant compounds were α -pinene (29.5 %), β -pinene (25.7%), and myrcene (13.0%), representing 68.2% of the HS volatiles on average. Abstract: The variability of volatiles isolated from twigs by the static headspace (HS) method in seven natural populations of Picea omorika from Serbia was investigated for the first time. In the overall chemical profile, monoterpenes strongly dominated hydrocarbons as the most volatile compounds (95.7%). The dominant compounds were α -pinene (29.5 %), β -pinene (25.7%) and myrcene (13.0%), totaling 68.2% of the volatiles on average. The following nine volatiles were found to be present in medium-to-high amounts (0.5-10%): tricyclene, camphene, α- phellandrene, δ -3-carene, p -cymene, β- phellandrene, terpinolene, ( E )-caryophyllene, and germacrene D. Out of the 78 volatiles detected, the six most abundant ( α -pinene, β -pinene, myrcene, δ -3-carene, β -phellandrene and camphene) were selected for principle component analysis (PCA) and cluster analyses (CA). PCA revealed a high degree of similarity between populations, while CA showed a degree of separation of two populations from the others. The presented results are in agreement with previous phytochemical and molecular analyses of this species that confirm high variability in both specialized metabolites and genetic markers. https://doi.org/10.2298/ABS200511038N Received: May 11, 2020; Revised: August 19, 2020; Accepted: August 24, 2020; Published online: August 27, 2020 How to cite this article: Nikolic B, Ljujic J, Bojovic S, Mitic Z, Rajcevic N, Tesevic V, Marin PD. Headspace volatiles isolated from twigs of Picea omorika from Serbia. Arch Biol Sci. 2020;72(3):445-52.

The Trametes versicolor laccase catalyzed oxidation of chalcone butein afforded four dimers of aurone sulfuretin (i.e. two regioisomeric pairs of diasteromers, 1-4) as the major products. The formation of the dimers was explained by a two step process involving the initial cyclization of butein into aurone sulfuretin, followed by the combination of two molecules of sulfuretin. The coupling process occurred between the 2,10-double bond of one molecule of sulfuretin and the (3′,4′) catechol group of the other to yield a dimeric structure. This was confirmed by the experiment involving the laccase catalyzed oxidation of sulfuretin yielding the same dimeric bisaurones. Compounds 1, 3 and 4, were isolated using semipreparative HPLC and characterized by the detailed analysis of the NMR, MS, IR, and UV-vis data. The structure of compound 2, isolated as a mixture containing ca. 25% of compound 1, was proposed by the comparison of 1H NMR data to compound 1 and by using LC-ESIMS analysis. The starting chalcone butein and the products of the biocatalytic transformation, aurone sulfuretin and sulfuretin dimers 1, 3 and 4, were evaluated for their cytotoxic and antioxidative properties in vitro using a healthy human fibroblast (MRC5) cell line. The biotransformation products showed lower cytotoxicity but higher antioxidative properties. The C. coggygria bark methanol extract rich in butein and sulfuretin was also biotransformed by laccase. The transformed extract exhibited significantly improved antioxidative activities. This is the peer-reviewed version of the article: M. Novakovic, T. Ilic-Tomic, V. Teševi, K. Simic, S. Ivanovic, S. Simic, I. Opsenica and J. Nikodinovic-Runic, New J. Chem., 2020, DOI: [https://doi.org/10.1039/D0NJ00758G] The published version: [https://cer.ihtm.bg.ac.rs/handle/123456789/3639]

Bat guano is an important source of microbial diversity in caves and can be a source of potential pathogens. Laemostenus (Pristonychus) punctatus is a guanophilic ground beetle species, which pygidial gland secretion exhibits action against pathogenic and other microbes. The distribution and diversity of microbes in bat guano from a karstic cave were determined in this study. Additionally, antimicrobial activity of the pygidial gland secretion of L. (P.) punctatus against guano-dwelling microbes was tested; minimal inhibitory concentration (MIC) and chemical composition of the secretion were analyzed. In total, 63 different bacterial species and 16 fungal morphotypes were isolated from guano samples by the cultivation method and confirmed using and phenotypic characterization and molecular identification. There was a difference in the composition of certain microorganisms between the sampling points (cave locations) and between the guano layers. The largest number of bacterial isolates belongs to the genera Lysinibacillus and Paenibacillus, while Pseudomonas species were highly abundant at the innermost sampling point. For the guanophilic fungi, the majority are ascomycetes, with Penicillium and Aspergillus as the most dominant genera. Meyerozyma guilliermondii was the only yeast species found in the guano samples. The most sensitive isolates were Enterococcus eurekensis (MIC 0.007 mg/mL) and Escherichia fergusonii (MIC 0.028 mg/mL). The most sensitive fungal isolates were M. guilliermondii, Penicillium expansum, and Trichoderma harzianum (MIC 0.15 mg/mL). This study opens a new possibility for better understanding of ecological relations between microorganisms and troglophilic ground beetles and for detailed investigations of morpho-anatomical aspects of pygidial glands.

The reliability of diarylheptanoids as chemotaxonomic markers at inter- and intraspecific levels has been investigated. Six diarylheptanoids were quantified in bark ethanol extracts of four Alnus spp. populations by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The populations described here as locus classicus Alnus glutinosa (I) and locus classicus Alnus incana (III) are clearly differentiated. Compared to population I, individuals in population III have higher extract yields and hirsutanonol-5-O-β-D-glucopyranoside content and lower contents of hirsutanonol, rubranoside A and oregonin. Individuals from neighboring populations of A. glutinosa (II) and A. incana (IV) share similar contents of hirsutanonol. All the studied populations exhibit a high intrapopulation variability of the selected diarylheptanoids; they have a heterogeneous chemotype and they partially overlap. The geographical proximity of populations II and IV increases their chemical similarity. Principal component analysis (PCA) clearly shows that the biggest dispersion of individuals lies within population IV. The reason for its heterogeneity might be its physical proximity to population II, i.e. the appearance of hybrids. Also, geometric morphometrics of leaves was performed as a screening criterion for spontaneous hybrids.

The contents of quercetin glycosides and neochlorogenic acid in fresh berries of Cornelian cherry and Common dogwood were assessed through LC-MS/MS analysis to determine chemical differentiation between two distant populations of each species. The aim of this study was to investigate the chemical variability of Cornus mas and Cornus sanguinea on the basis of the content of six selected phenolic compounds in fruit extracts. Fruits were sampled at the time of full ripening, mid-September, from two localities that differed in terms of orographic and environmental conditions. Fresh fruit extracts were analyzed by LC-MS/MS to determine the presence and contents of neochlorogenic acid, quercitrin, isoquercetin, hyperoside, rutoside and querciturone. While all of the tested compounds were detected in both species, their content was several times higher in C. sanguinea. Locality-dependent differences were tested by appropriate statistical tests, which confirmed that the contents of all compounds significantly differed in C. mas, whereas the content of only one compound differed in C. sanguinea. Differences between populations were also revealed by PCA analyses. Both species showed clear separation of populations based on the analyzed phenolic compounds. The results showed that a specific environment can significantly affect the contents of certain secondary metabolites, as in the case of C. mas, and/or that the synthesis of these compounds can be to a lesser extent influenced by external factors, as in the case of C. sanguinea.

Seven derivatives of pentacyclic triterpene acids (1-7) were isolated from the bark of Alnus viridis ssp. viridis using a combination of column chromatography and semipreparative HPLC. Compounds 1-3, 6, and 7 were determined to be new after spectroscopic data interpretation and were assigned as 27-hydroxyalphitolic acid derivatives (1-3), a 27-hydroxybetulinic acid derivative (6), and a 3-epi-maslinic acid derivative (7), respectively. Pentacyclic triterpenoids with a C-27 hydroxymethyl group have been found in species of the genus Alnus for the first time. These compounds were subjected to cytotoxicity testing against a number of canter cell lines. Also, selected pentacyclic triterpenoids were selected as potential inhibitors of topoisomerases I and Ha for an in silico investigation. Supplementary material: [http://cherry.chem.bg.ac.rs/handle/123456789/3088]

The variations of metabolic profile of the latex of wild-growing Euphorbia palustris was carried out using multivariate analysis of NMR spectral data. One population was infected with fungi Fusarium sporotrichioides, Fusarium proliferatum and Alternaria alternata, while the other consisted of healthy plant species. The non-polar metabolites of latex extracts such as benzoyl ingenol-laurate, amyrin decadienoate esters, cis-1,4-polyisoprene, and 24-methylenecycloartanol were identified using H-1 and 2D NMR spectra. Principal component analysis of H-1 NMR data provided a clear discrimination between the latex of infected and healthy plants. Minimum inhibitory concentration and minimum fungicidal concentration values of the latex extracts of healthy and infected plants were determined. The latex of infected plants was found to contain higher levels of benzoyl ingenol-laurate and 24-methylenecycloartanol, of which concentrations were strongly correlated with the antifungal activities of the latex. (C) 2016 Elsevier Ltd. All rights reserved. Supplementary material: [http://cherry.chem.bg.ac.rs/handle/123456789/3564]

Metabolomics generate a profile of small molecules from plant extracts, which could be directly responsible for bioactivity effects. Using dry-column flash chromatography enabled a rapid and inexpensive method for the very efficient separation of plant extract with a high resolution. This separation method coupled to NMR and FTIR-based metabolomics is applied to identify bioactive natural products. OPLS multivariate analysis method, was used for correlation the chemical composition of the plant extracts, Amphoricarpos autariatus, with the results of cytotoxic activity against Human cervical adenocarcinoma cell line (HeLa) and epithelial lung cancer cell line (A549). In this way, the highest contribution to the cytotoxic activity was recorded for the guaianolide sesquiterpene lactones named amphoricarpolides. The compounds indicated as bioactive after metabolomics analysis were tested, and their cytotoxic activity were confirmed. Supplementary material: [http://cherry.chem.bg.ac.rs/handle/123456789/3349]

Seven new bisbibenzyls (1-7) were isolated from the methanol extract of the liverwort Lunularia cruciata along with one previously known bibenzyl and five known bisbibenzyls. The structures of compounds 1-7 were elucidated on the basis of the spectroscopic data. These newly isolated bisbibenzyls may be divided into two groups, the acyclic bisbibenzyls, perrottetins (1-3), and the cyclic analogues, riccardins (4-7). Besides standard perrottetin and riccardin structures (1 and 4, respectively), they contain phenanthrene (3 and 5), dihydrophenanthrene (2), and quinone moieties (6 and 7), rarely found in natural products. The new compounds 3 and 5, as well as the known riccardin G, exhibited cytotoxic activity against the A549 lung cancer cell line with IC 50 values of 5.0, 5.0, and 2.5 μM, respectively. Supplementary material: [http://cherry.chem.bg.ac.rs/handle/123456789/3097]

Volatiles of Thymus serpyllum L. were obtained by hydrodistillation (HD), simultaneous hydrodistillation, and extraction (SDE) and static headspace gas chromatography-mass spectrometry analysis (head space [HS]), respectively. Monoterpenes were the most dominant in all 3 techniques (84.8%-94.2%). Essential oil profiles obtained by HD and SDE were similar, with oxygenated monoterpenes being the most abundant (up to 75.4%). In HS volatiles of T. serpyllum monoterpene hydrocarbons strongly dominated (94.2%). The main compounds were alpha-terpinyl acetate (HD and SDE) and myrcene (HS).

The presence of n -alkanes, free alcohols and free acids in leaf epicuticular wax extracts of 22 samples of 11 Euphorbia L. species belonging to the sections Paralias , Esula , Myrsiniteae and Helioscopia , 10 of which were never examined before, were analyzed by gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS), and n -alkane C27 was detected as the principal component of leaf epicuticular waxes in the majority of the examined species, while the most abundant free alcohol was C26. Three Euphorbia species belonging to section Helioscopia were characterized by a predominance of alcohol C28. Free acid (C16) was the major component in 21 investigated samples. The usefulness of n -alkanes and free alcohols and free acids as potential chemotaxonomic markers is briefly discussed. https://doi.org/10.2298/ABS180608041K Received: June 8, 2018; Revised: August 31, 2018; Accepted: September 12, 2018; Published online: September 21, 2018 How to cite this article: Krstic G, Aljancic I, Stankovic J, Cvetkovic M, Marin P, Janackovic P, Tesevic V. Leaf epicuticular waxes of eleven Euphorbia species (Euphorbiaceae) from the Central Balkans: Impact on chemotaxonomy. Arch Biol Sci. 2019;71(1):21-6.

This study was performed to determine the main antibacterial compounds of the essential oil (ЕО) of saltmarsh plant Artemisia santonicum (Asteraceae). The combination of HPTLC and direct bioautography was used for the activity guided isolation of isogeranic acid as the main antibacterial constituent with remarkable antimicrobial activity, although it was the minor component of the EO, present only in 0.2 %, as calculated from GC/FID. Its structure was determined by 1D- and 2D-NMR and GC–MS techniques. Antibacterial activity of isogeranic acid against all tested bacteria was significantly higher than EO and even than both controls streptomycin and ampicillin. In further investigation of antibiofilm and antiquorum sensing activity EO exhibited the best inhibition of the biofilm formation at 1/8 minimal inhibitory concentration (MIC) and isogeranic acid at 1/2 MIC. Both EO and isogeranic acid possessed pyocyanin inhibitory activity showing the reduction of pigment at 60.6 and 62.8 %, respectively, at 1/2 MIC concentrations. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. 172053, Grant no. 173032 and Grant no. TR 31089]

Two sets of structurally different jatrophanes (1-11 and 13-16), jatrophane 12, and latex extract of 2 Euphorbia species (17 and 18) were tested for in vitro protective effect against chromosome aberrations in peripheral human lymphocytes using the cytokinesis-block micronucleus (CBMN) assay. Jatrophanes 1-6 in minimal doses of 1 µg/mL prominently decreased micronuclei (MN) frequency in the range 44.86% to 34.29% and manifested considerable protective effect. From the other set of jatrophanes, 13 in the same minimal dose notably decreased MN frequency by 31.05%, while extracts 17 and 18 at a concentration of 4 µg/mL remarkably decreased the frequency of MN by 37.94% and 36.12%, respectively. Jatrophanes 12, 14, and 16 showed moderate protection, while 7-11 and 15 were less active than positive control. The structure-activity relationship (SAR) studies of the tested jatrophanes (1-16) indicated the favorable position of benzoate at C-8 or C-9 (3, 4, and 13) and a preference of isobutanoyloxy group at C-3 (1-3) rather than propanoyloxy at the same position (4-6) for pronounced protective effect on human lymphocytes DNA. In a previous SAR study on 11 jatrophanes (1, 3-8, and 13-16), the same structural features in 3, 4, and 13 influenced powerful inhibition of P-gp, while growth inhibition of cancer cells was more than doubled in 1 (isobutanoyloxy group at C-3) compared to 6 (propanoyloxy at C-3).

Species of the Lamiaceae family have enjoyed a rich tradition of use for flavoring, food preservation, and medicinal purposes, due to their curative and preventive properties. Cultivated winter savory ( Satureja montana L.), sage ( Salvia officinalis L.) and hyssop ( Hyssopus officinalis L.) are produced for seed, herb, and essential oil. Dominant compounds in S. montana essential oil were carvacrol (43.2%) and thymol (28.4%), while cis -thujone (27.1%) and camphor (19.3%), followed by trans -thujone and 1,8-cineole were the major compounds in S. officinalis essential oil. As for H. officinalis essential oil, cis - and trans -pinocamphone (41.1% and 20.5%, respectively) were the most abundant compounds, followed by β-pinene. The highest antimicrobial properties, as well as antioxidant capacity, showed S. montana essential oil compared to other tested essential oils. Furthermore, H. officinalis essential oils showed higher antioxidant activity than that of S. officinalis . The aim of this investigation was to determine the composition and bioactivity of essential oils of mentioned varieties. Presented results show that S. montana essential oil could be proposed as a valuable source of natural preservatives.

This work aimed to evaluate the content of selected phenolic natural products in the wine samples made of three new Serbian Cabernet Franc clones ( Nos. 02, 010 and 012, respectively) and mother vine ( used as the relevant standard) during the period 2008-2012. Compared with all other wine samples, the Cabernet Franc wine of the clone No. 010 was found to have the highest total content of polyphenolics ( 1.85 +/- 0.02 g/L) and anthocyanins ( 178.55 +/- 3.75 mg/L). In addition, its Folin-Ciocalteu index ( 36.86 +/- 0.12) stood out among the examined samples. Finally, the same wine was enriched with ellagic and gallic acids ( 3.44 +/- 0.29 and 27.46 +/- 0.21 mg/L, respectively), catechin ( 135.16 +/- 6.47 mg/L) and epicatechin ( 51.33 +/- 2.33 mg/L), the natural products known to exert significant lipid-lowering effects. Taken all together, the clone No. 010 developed in Serbia may offer new Cabernet Franc wine with geographical indication. [GRAPHICS] . Supplementary material: [http://cherry.chem.bg.ac.rs/handle/123456789/3652]

Methanol extracts of selected species of flowering plants Anthemis cotula, A. ruthenica, Centaurea dubia (Asteraceae), Ajuga genevensis, A. chamaepitys, A. reptans, Micromeria albanica, M. cristata, M. dalmatica, M. juliana, Thymus tosevii (Lamiaceae) and conifers - Abies alba, Picea omorika, Pinus heldreichii (Pinaceae) and Taxus baccata (Taxaceae), as well as diethyl ether extracts of ten species Anthemis cotula, A. ruthenica, Centaurea dubia, Ajuga genevensis, A. chamaepitys, A. reptans, Micromeria albanica, M. cristata, M. dalmatica and M. juliana from two flowering plant families (Asteraceae and Lamiaceae) were tested for general bioactivity using brine shrimp (Artemia salina) lethality test. Lethal concentration (LC50) and 95% confidence intervals were determined by computer program LdP line. Out of fifteen tested methanol extracts, three possessed cytotoxic effect. Taxus baccata methanol extract showed the highest effect (LC50 = 18.60 μg/ml), while Thymus tosevii methanol extract expressed the lowest (LC50 = 842.50 μg/ml). All other analyzed species did not express significant cytotoxicity. Also, diethyl ether extracts of all tested species did not show significant cytotoxicity. The obtained results for methanol extracts which show certain cytotoxic effect could be guide for further phytochemical and pharmacological investigations. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. 173029]

Horse chestnut (Aesculus hippocastanum L.) is a deciduous tree which is widely used on urban green spaces in Serbia. Also, the seed of horse chestnut is an important raw material in pharmaceutical industry, owing to its medicinal properties. In this study, the content of fatty acids, flavonoids, aescin and heavy metals in seeds of 11 horse chestnut genotypes growing in urban conditions was investigated. A total of 16 fatty acids were identified. Unsaturated fatty acids in seed samples were the most abundat. Among the saturated fatty acids, palmitic acid was the dominant one, with a total average content of 8.12%. Oleic acid, with the total average content of 53.46%, was the most abundant unsaturated fatty acid. The results of this study indicated the presence of the following flavonoids: quercetin, kaempferol and rutin. Rutin was the most abundant, averagely contributing with 6.071 μg/g to the total quantified polyphenolics in all samples. The total aescin content ranged from 0.82% to 4.16%. Fruits of park and street trees from small- and middle-sized cities can be used in pharmaceutical industry as raw material. Differences between different urban site categories and localities were detected, which indicates the biochemical plasticity of the seeds to urban morphology. Supplementary material: [http://cherry.chem.bg.ac.rs/handle/123456789/3807]

Consumer acceptance of plum is based, primarily, on the corresponding sensorycharacteristics of the fruits: appearance, colour, firmness, taste and aroma. If notconsumed immediately after harvesting, the aforementioned characteristics of plumwill change. Still, those changes are less expressed when fruits are kept in coldstorage, at low temperatures. However, during the transport and storage of plum atroom temperature, which is often the case in local market plum sale, intensechanges of certain characteristics of the fruit happens as well as a significantshortening of shelf life. The most expressive is firmness change where fruitsbecome completely soft after seven days. Appearance and colour of the fruit areless susceptible to change. Since plum taste and odour depend on sugar and acidcontent, i.e., sugar-acid ratio and the content of certain volatile aromaticcomponents, changes of their contents have been examined in some importantplum cultivars in Serbia after seven-day storage at the temperature 20±1 °C. Sugaracidratio that determines sweetness, i.e., fruit acidity, has not significantly changedduring thе storage period. Among 10 examined aromatic components, the mostsignificant changes have been observed in 2-E-hexenal, which gives a distinctivegreen note aroma and nonanal, characteristic plum-like scent. During the seven-daystorage of picked plums, the content of 2-E-hexenal decreased while the content ofnonanal increased.

The n-alkane composition in the leaf cuticular waxes of natural populations of Bosnian pine (Pinus heldreichii), Austrian pine (P.nigra), and Macedonian pine (P.peuce) was compared for the first time. The range of n-alkanes was wider in P.nigra (C-16-C-33) than in P.heldreichii and P.peuce (C-18-C-33). Species also diverged in abundance and range of dominant n-alkanes (P. heldreichii: C-23, C-27, and C-25; P.nigra: C-25, C-27, C-29, and C-23; P.peuce: C-29, C-25, C-27, and C-23). Multivariate statistical analyses (PCA, DA, and CA) generally pointed out separation of populations of P.nigra from populations of P.heldreichii and P.peuce (which were, to a greater or lesser extent, separated too). However, position of these species on the basis of n-alkane composition was in accordance neither with infrageneric classification nor with recent molecular and terpene investigations. This is the peer-reviewed version of the article: Nikolić, B.; Mitić, Z. S.; Tešević, V.; Đorđević, I.; Todosijević, M.; Bojović, S.; Marin, P. D. Chemotaxonomic Considerations of the N-Alkane Composition in Pinus Heldreichii, P.Nigra, and P.Peuce. Chemistry & Biodiversity 2018, 15 (9), e1800161. [https://doi.org/10.1002/cbdv.201800161]

A number of potentially useful compounds with various biological activities can be obtained from plant metabolites by microorganisms. In present research, essential oil of Calamintha glandulosa was subjected to the biotransformation process by mycromicete Aspergillus niger. GC/MS analyses of the original and biotransformed essential oil have revealed six(1-6) biotransformed compounds from one of the main component-limonene and one biotransformed product (7) originating from the less abundant component-thymol. Piperitone epoxide and piperitenone epoxide, the most abundant components of the original oil, were not biotransformed but gave three artefacts (A1, A2, B1) due to the acidic conditions formed in A. niger medium. Compounds 5, 6 as well as 7 have not been previously found as a biotransformation products of limonene (5 and 6) and thymol (7), respectively. New products (5, 6, and 7) together with the artefacts, were isolated by silica gel column chromatography followed by preparative silica gel thin layer chromatography, and their structures were established by a combination of 1D and 2D NMR spectra and GC/MS data. Biotransformation has one more confirmed as useful method, complementary to synthesis, for obtaining different type of molecules. Supplementary data: [https://cer.ihtm.bg.ac.rs/handle/123456789/4448]

The essential oil of Thymus algeriensis was analyzed as a potential preservative in soft cheese. We developed a novel method to test the preserving properties of essential oil in soft cheese. Contamination incidence of Penicillium aurantiogriseum was absent after 30 d of storage at 4 degrees C with 25 mu L of essential oil added. The antimicrobial activity was tested against 8 bacteria and 8 fungi. Thymus algeriensis oil showed inhibitory activity against tested bacteria at 0.03 to 0.09 mg/mL, and bactericidal activity was achieved at 0.05 to 0.15 mg/mL. For antifungal activity, minimum inhibitory concentrations ranged between 0.01 and 0.04 mg/mL and minimum fungicidal concentrations between 0.01 and 0.04 mg/mL. Furthermore, the oil was also screened for antiradical activity using the 1,1-diphenyl-2-picrylhydrazyl assay. The results showed that the oil was active and achieved half-maximal inhibitory activity at 0.132 mg/mL. We used gas chromatography, gas chromatography-mass spectrometry, and nuclear magnetic resonance spectrometry to investigate the volatile compounds from the oil. Carvacrol was identified as the main compound in the oil, represented by 80.9% of the total constituents, followed by p-cymene (7.7%).

In a last few decades mushrooms are increasingly attracting attention as functional food and sources of biologically active compounds. Several Trametes species have been used for centuries in traditional medicine of East Asia cultures, but only T. versicolor was studied sufficiently while there are less substantial data about medicinal properties of other species. Trametes versicolor, T. hirsuta and T. gibbosa were the species tested for biological activities. Antifungal potentials of extracts were assessed for clinical strains of selected Candida and Aspergillus species. ABTS and FRAP assays were used to evaluate antioxidant capacities of studied extracts. Cytotoxic activity was determined against human cervix and lung adenocarcinoma and colon carcinoma cell lines. Antineurodegenerative activity was assessed by determining the rate of acetylcholinesterase and tyrosinase activity. The presence of metabolites in extracts of mycelia and basidiocarps of studied Trametes species was analyzed by 1H NMR spectroscopy. Studied extracts showed low antifungal potential in comparison with ketoconazole. Basidiocarp extracts were more effective ABTS+ scavengers and Fe2+ reducers than mycelium ones but less effective in comparison with L-ascorbic acid. Results showed that mycelium extracts had stronger cytotoxic effects against three cancer cell lines than basidiocarp ones, and that cervix adenocarcinoma cells were the most sensitive to the extracts and commercial cytostatics. T. versicolor mycelium extract was the most effective inhibitor of acetylcholinesterase activity but double weaker than galantamine, and T. gibbosa mycelium extract was significantly better inhibitor of tyrosinase activity than kojic acid for 40.9%. Chemical analysis indicated strong synergistic action of triterpenes, sugars and polyphenols in applied assays. The results suggest that tested Trametes species have significant medicinal potentials which could be attributed to antioxidative and cytotoxic activity. Additionally both, basidiocarps and mycelia extracts can strongly inhibit activity of acetylcholinesterase and tyrosinase.

Diarylheptanoids are a group of secondary metabolites widely distributed in the Betulaceae family and characteristic for Alnus species. In this study, the chemotaxonomic power of diarylheptanoids, such as hirsutanonol-5-O-β-d-glucopyranoside, rubranoside A, oregonin, platyphylloside, alnuside A and hirsutanonol, has been investigated in combination with principal component analysis (PCA) for differentiation of Alnus species. Concentrations of six diarylheptanoids in the bark extracts of two natural populations of Alnus glutinosa (black alder) and Alnus incana (gray alder) were determined by ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). PCA clearly shows the separation of three groups. Populations I (A. glutinosa) and II (A. incana) both consisted of individuals of the corresponding species. Four individuals from both sampled populations formed a separate group (population III), which possibly represents a hybrid group. Accordingly, diarylheptanoids may serve in combination with PCA as chemotaxonomic markers at the species level, which may also reveal hybrid species.

The composition of the essential oil (EO) obtained by hydrodistillation from the leaves of Thymus capitatus Hoff. et Link. growing wild in Sythonia (Greece) was analysed by GC-FID, GC-MS and quantitative H-1 NMR spectroscopy. Thirty nine compounds were identified, representing approximately 99.0 % of the oil. Major component determined by GC-FID was carvacrol (81.8 %), followed by linalool (3.5 %) and (E)-caryophyllenc (3.5 %). Abundance of carvacrol determined by quantitative H-1 NMR spectroscopy was 74.9 % and was in reasonable agreement with GC-FID result. The oil was tested for the antioxidant activity in DPPH assay and showed a dose dependent free radical scavenging activity with EC50 value of 1.03 mu g/ml. Antimicrobial effect of the EO was tested against Gram negative, Gram positive bacteria, and fungi. The tested EO showed significant activity against fungi, especially S. cerevisiae. The most sensitive bacteria were E. coil ATCC 25922, E. fecalis and S. epidermidis. Human pathogen Burkholderia cepacia, ATCC 25416 was used in the investigation of T capitatus for the fast time.

New aurone epoxide, 2,10-oxy-10-methoxysulfuretin (14), and new auronolignan (15), named cotinignan A, were isolated by silica gel column and semipreparative HPLC chromatography from the methylene chloride/methanol extract of Cotinus coggygria Scop. heartwood. In addition, thirteen known secondary metabolites namely sulfuretin, 2,3-trans-fustin, fisetin, butin, butein, taxifolin, eriodictyol, 3',5,5',7–tetrahydroxyflavanone, 3',4',7-trihydroxyflavone, 3-O-methyl-2,3-trans-fustin, 3-O-galloyl-2,3-trans-fustin, β-resorcylic acid and 3-O-β-sitosterol glucoside were isolated as well. Their structures were elucidated by 1D and 2D NMR, HR-ESI-MS, IR and UV. Ten out of eleven isolated flavonoids possess 7, 3' and 4' hydroxy groups. These structural features could be considered as chemotaxonomic characteristic of flavonoids from C. coggygria. Cotinignan A (15) represents new subclass of secondary metabolites - auronolignans. (Figure presented.). Supplementary material: [https://cer.ihtm.bg.ac.rs/handle/123456789/4447]

In the light of growing interest in discovering new sources of natural antimicrobial agents, we extracted and tested the efficiency of julid defensive secretions in comparison with commercially used antibiotic and antimycotics. This work involved chemical characterization of the defensive secretions of Cylindroiulus boleti (C. L. Koch, 1847), Megaphyllum bosniense (Verhoeff, 1897) and M.unilineatum (C. L. Koch, 1838), as well as in vitro evaluation of their antimicrobial activity against 11 bacteria, one yeast and eight filamentous fungi. Compounds of the analysed defensive secretions included p-benzoquinones, alkyl esters of fatty acids and ketones. Ketones were recorded for the first time in the order Julida, and they were detected in secretions of both Megaphyllum species. All three analysed defensive secretions showed antibacterial and antifungal potential against all of the tested pathogens. Staphylococcus aureus proved to be the most sensitive bacterial strain to all analysed secretions. Also, defensive secretion of M.unilineatum showed significant antibacterial potential against Bacillus subtilis. The most resistant bacterial strains in this study were Escherichia coli, Erwinia persicina and Pseudomonas syringae pv. tomato. Analysed defensive secretions achieved the strongest antifungal activity against Aspergillus parasiticus (secretions of all three millipede species), Penicillium griseofulvum (secretions of C.boleti and M.bosniense) and Cladosporium oxysporum (secretion of M.unilineatum). The most resistant mycromycetes were A.niger (to all tested defensive secretions), A.flavus (to secretion of M.unilineatum) and P.lanosum (to secretions of C.boleti and M.bosniense). Our results showed a generally lower level of activity compared to antibiotic and a significantly higher level compared to antimycotics. The results of this study elucidate and open opportunities for further research in the field of millipede chemical ecology. Supplementary material: [http://cherry.chem.bg.ac.rs/handle/123456789/3238]

Seven previously undescribed jatrophane diterpenoids, nicaeenin A-G, with eight known jatrophane diterpenoids, namely euphodendrophanes A-C, F, N, O, Q S, were isolated from latex of Euphorbia nicaeensis collected in Serbia. The chemical structures of the compounds were determined by spectro-scopic analysis including 1D and 2D NMR and HRESIMS experiments. All but one of the previously undescribed jatrophanes, showed significant potential to inhibit P-glycoprotein (P-gp) activity in two MDR cancer cells (NCI-H460/12 and DLD1-TxR). The most powerful were nicaeenin F and nicaeenin G. Moreover nicaeenin G significantly stronger sensitized NCI-H460/R cells to DOX than Dex-VER. (C) 2018 Elsevier Ltd. All rights reserved. Supplementary material: [http://cherry.chem.bg.ac.rs/handle/123456789/3234]

Ethnopharmacological relevance: Meadowsweet (Filipendula ulmaria (L.) Maxim.) and dropwort (Filipendula vulgaris Moench) are herbaceous perennials employed in folk medicine for their antirheumatic, antipyretic and antiulcer properties. Aim of the study: To assess ethnomedicinal claims through investigation of antioxidant, anti-inflammatory and gastroprotective effects of F. ulmaria and F. vulgaris lyophilized flower infusions (LFIs) as well as the F. vulgaris isolated flavonoids spiraeoside, kaempferol 4'-O-glucoside, astragalin 2'-O-gallate, mixture of hyperoside 2'-O-gallate and isoquercitrin 2'-O-gallate, and a tannin tellimagrandin IL Materials and methods: Free radical scavenging activity of the tested samples was determined by examining their ability to neutralize DPPH and OH radicals in vitro, whereas reducing properties were assessed in Ferric Reducing Antioxidant Power (FRAP) assay. Anti-inflammatory activity was studied ex vivo in human platelets by monitoring the effect on eicosanoid biosynthesis.. Gastroprotective action was estimated in animal model of acute gastric injury induced by ethanol. Results: LFIs and spiraeoside exerted activities comparable to those of positive control in DPPH-radical scavenging and FRAP antioxidant assays, whereas notable hydroxyl radical scavenging ability was demonstrated only for spiraeoside (IC50 = 5.1 mu g/mL). Among tested samples, astragalin 2 ''-O-gallate (IC50 = 141.1 mu g/mL) and spiraeoside (IC50 = 4.69 mu g/mL) the most markedly inhibited production of pro -inflammatory prostaglandin E-2 and 12(S)-hydroxy-(5Z,8Z,10E,14Z)-eicosatetraenoic acid in human platelets, respectively. Examination of LFIs (100-300 mg/kg, p.o.) gastroprotective action in rats revealed their capacity to preserve mucosal integrity. In addition, spiraeoside (50 mg/kg, p.o.) and tellimagrandin II (40 mg/kg, p.o.) showed ulcer preventive ability. Conclusion: Current study supports documented traditional use of investigated herbs and indicates that flavonoid and tannin components are partially responsible for the demonstrated pharmacological activities. This is peer-reviewed version of the following article: Samardžić, S.; Arsenijević, J.; Božić, D.; Milenković, M.; Tešević, V.; Maksimović, Z. Antioxidant, Anti-Inflammatory and Gastroprotective Activity of Filipendula Ulmaria (L.) Maxim. and Filipendula Vulgaris Moench. Journal of Ethnopharmacology 2018, 213, 132–137. [https://doi.org/10.1016/j.jep.2017.11.013] Supplementary material: [http://cherry.chem.bg.ac.rs/handle/123456789/3154]

In this study, we compared the compositions, antimicrobial and potential antioxidant activities of essential oils of two Thymus endemic, wild-growing species - T. malyi Ronninger and T. lykae Degen et Jay. No information about its composition and biological activities of oils has been reported to the present day. Forty three compounds were identified from T. malyi and major component was a-pinene (26.4%). In the case of T. lykae, forty four compounds were identified, with geranyl acetate (35.1%) as the main component. Antioxidant activities of the oils were evaluated using DPPH assay. The antimicrobial effect of essential oils were tested against Gram-negative, Gram-positive bacteria, and two fungi. Results indicated higher antibacterial activity of the essential oils of T. lykae in comparison with T. malyi, while for antifungal activity, it was reverse. The essential oils from T. malyi and T. lykae possess antioxidant and antimicrobial activities and could be used as a potential source of natural antioxidants for the food industry.

The composition of the epicuticular n-alkanes isolated from the leaves of ten populations of Juniperus communis L. var. saxatilis PALLAS from central (continental) and western (coastal) areas of the Balkan Peninsula was characterized by GC-FID and GC/MS analyses. In the leaf waxes, 14 n-alkane homologues with chain-lengths ranging from C-22 to C-35 were identified. All samples were dominated by n-tritriacontane (C-33), but differences in two other dominant n-alkanes allowed separating the coastal from the continental populations. Several statistical methods (ANOVA, principal component, discriminant, and cluster analyses as well as the Mantel test) were deployed to analyze the diversity and variability of the epicuticular-leaf-n-alkane patterns of the ten natural populations of J. communis var. saxatilis and their relation to different geographic and bioclimatic parameters. Cluster analysis showed a high correlation of the leaf-n-alkane patterns with the geographical distribution of the investigated samples, differentiating the coastal from the continental populations of this taxon. Several bioclimatic parameters related to aridity were highly correlated with this differentiation.

The antifungal potential of the pygidial gland secretion of the troglophilic ground beetle Laemostenus punctatus from a cave in Southeastern Serbia against cave-dwelling micromycetes, isolated from the same habitat, has been investigated. Eleven collected samples were analyzed and 32 isolates of cave-dwelling fungi were documented. A total of 14 fungal species were identified as members of the genera Aspergillus, Penicillium, Alternaria, Cladosporium, Rhizopus, Trichoderma, Arthrinium, Aureobasidium, Epicoccum, Talaromyces, and Fusarium. Five isolates were selected for testing the antifungal activity of the pygidial gland secretion : Talaromyces duclauxi, Aspergillus brunneouniseriatus, Penicillium sp., Rhizopus stolonifer, and Trichoderma viride. The microdilution method has been applied to detect minimal inhibitory concentrations (MICs) and minimal fungicidal concentrations (MFCs). The most sensitive isolate was Penicillium sp., while the other isolates demonstrated a high level of resistance to the tested agent. L. punctatus has developed a special mechanism of producing specific compounds that act synergistically within the secretion mixture, which are responsible for the antifungal action against pathogens from the cave. The results open opportunities for further research in the field of ground beetle defense against pathogens, which could have an important application in human medicine, in addition to the environmental impact, primarily.

Comparative analysis of terpene diversity and differentiation of relict pines Pinus heldreichii, P.nigra, and P. peuce from the central Balkans was performed at the population level. Multivariate statistical analyses showed that the composition of needle terpenes reflects clear divergence among the pine species from different subgenera: P. peuce (subgenus Strobus) vs. P. nigra and P. heldreichii (subgenus Pinus). In addition, despite the described morphological similarities and the fact that P. nigra and P. heldreichii may spontaneously hybridize, our results indicated differentiation of their populations naturally growing in the same area. In accordance with recently proposed concept of flavonic evolution' in the genus Pinus, we assumed that the terpene profile of soft pine P. peuce, defined by high amounts of six monoterpenes, is more basal than those of hard pines P. nigra and P. heldreichii, which were characterized by high content levels of mainly sesquiterpenes. In order to establish precise positions of P. heldreichii, P. nigra and P. peuce within the taxonomic and phylogenetic tree, as well as develop suitable conservation strategies and future breeding efforts, it is necessary to perform additional morphological, biochemical, and genetic studies. This is the peer-reviewed version of the following article: (1) Mitić, Z. S.; Nikolić, B. M.; Ristić, M. S.; Tešević, V. V.; Bojović, S. R.; Marin, P. D. Terpenes as Useful Markers in Differentiation of Natural Populations of Relict Pines Pinus Heldreichii, P. Nigra, and P. Peuce. Chemistry and Biodiversity 2017, 14 (8). [https://doi.org/10.1002/cbdv.201700093]

The essential oil, different extracts, and isolated compounds of Angelica pancicii Vandas (Apiaceae) were investigated for the first time. The GC-FID and GC-MS analyses revealed sesquiterpenoids as the main constituents of A. pancicii essential oil of aerial parts with bornyl acetate (8.08%), n-octanol (5.82%), kessane (4.26%), and beta-selinene (4.26%) as the main constituents. Analysis of methanol extracts, using an HPLC-DAD/ESI-ToF-MS system, showed a total of 52 compounds in the aerial parts and 53 in the roots, indicating coumarins as the main constituents. In addition, new chromone (1) and six known furanocoumarins (2-7) were isolated from the roots and structurally elucidated by combined spectroscopic methods. The aerial part extracts exhibited higher polyphenolic contents and antioxidant activity evaluated by three radical scavenging assays. Using a microwell dilution method, the strongest antibacterial activity profiles were determined for ethanol and methanol root extracts (minimum bactericidal concentrations (MBCs) = 0.25-3.00 mg/mL), which were comparable to the activity of streptomycin (MBCs = 0.34-1.24 mg/mL), while the strongest antibacterial compound of A. pancicii was oxypeucedanin hydrate (MBCs = 0.50-8.00 mg/mL). Antifungal potential was in moderate extent, and the highest activity was obtained for root methanol extract (minimum fungicidal concentrations (MFCs) = 4.00-14.00 mg/mL). Tested sub-minimum inhibitory concentrations (subMICs) of the extracts and isolated compounds inhibited selected Pseudomonasaeruginosa PAO1 virulence determinants. The most reduced growth of P. aeruginosa colony was in the presence of isolated oxypeucedanin. Ethanol (17.36-46.98%) and methanol (34.54-52.43%) root extracts showed higher anti-biofilm activity compared to streptomycin (49.40-88.36%) and ampicillin (56.46-92.16%). This a peer-reviewed version of the article: [https://doi.org/10.1021/acs.jafc.7b04202] [http://cer.ihtm.bg.ac.rs/handle/123456789/2110]

Brunfelsia genus is traditionally utilized in popular medicine due to its antibacterial and antifungal properties to name but a few. However, studies on the antimicrobial activity of Brunfelsia uniflora flower oleoresin have not been found yet. This study aimed to evaluate the chemical composition and antimicrobial activity of B. uniflora flower oleoresin obtained by supercritical carbon dioxide. Oleoresin from the plant dried flowers was obtained by carbon dioxide, and the chemical composition was analyzed by gas chromatographic-mass spectrometry. The minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and minimum fungicidal concentration (MFC) of this oleoresin for seven bacteria and eight fungi were determined using 96-well microtiter plates. The oleoresin MBC for Bacillus cereus, Enterobacter cloacae, Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella enterica, and Staphylococcus aureus ranged from 0.01 to 0.08 mg/mL, whereas the controls streptomycin and ampicillin varied from 0.1 and 0.5 mg/ mL. The oleoresin MFC for Aspergillus fumigatus, Aspergillus niger, Aspergillus ochraceus, Aspergillus versicolor, Penicillium funiculosum, Penicillium ochrochloron, Penicillium verrucosum var. cyclopium, and Trichoderma viride varied from 0.01 to 0.08 mg/mL, whereas the controls bifonazole and ketoconazole ranged from 0.2 to 3.5 mg/ mL. The oleoresin obtained by supercritical carbon dioxide presented bacteriostatic, bactericidal, fungistatic, and fungicidal activities that were higher than the positive controls streptomycin, ampicillin, bifonazole, and ketoconazole. The high antimicrobial activity was related to the high content of (E, E)-geranyllinalool that composes 21.0% of the oleoresin and a possible synergic action with fatty acid esters that made up 50.5% of the oleoresin. The oleoresin antimicrobial activity against common multiresistant bacteria in severe infectious processes as P. aeruginosa or against toxin-producing fungi such as P. ochrochloron or fungi that are difficult to control such as T. viride suggests the development of promising applications of this product in the food, farming, livestock, and pharmaceutical industry.

Analysis of composition of sesquiterpene lactone fraction of leaf cuticular neutral lipids of Amphoricarpos complex from two different localities in north Montenegro, i.e., canyon of river Tara (A. autariatus ssp. autariatus) and mountain Zeletin (A. autariatus ssp. bertisceus) afforded sesquiterpene lactones with guaianolide skeletons (17 compounds), so called amphoricarpolides, typical for this genus. Nine of them, 9-17, were new compounds, and their structures were elucidated by detailed analyses of IR, NMR, and MS data.

Nine diarylheptanoids, 1-9, catechin (11), and a phenolic glucoside, 10, were isolated from the bark of green alder (Alnus viridis). Four of the isolated compounds, i.e., 2, 5, 8, 10, are new. The structures of 1-11 were determined on the basis of spectroscopic data. All isolated compounds were evaluated for their in vitro protective effects on chromosome aberrations in peripheral human lymphocytes using cytokinesis-block micronucleus (CBMN) assay. Almost all of them exerted a pronounced effect of decreasing DNA damage of human lymphocytes, acting stronger than the known synthetic protector amifostine.

The defensive secretions of two blaniulid millipedes, Nopoiulus kochii and Cibiniulus phlepsii, were characterized by GC-FID and GC/MS analyses, which showed the presence of a complex mixture of benzoquinones, hydroquinones, and oleates. Altogether, 13 compounds were identified. The major compound in the secretions of both analyzed species was 2-methyl-1,4-benzoquinone (toluquinone). The second major constituent in the N. kochii secretion was 2-methyl-3,4-(methylenedioxy)phenol, while in that of C. phlepsii, it was 2-methoxy-3-methyl-1,4-benzoquinone. The defensive secretion of N. kochii also showed a high content of hydroquinones (13.5%) in comparison to that of C. phlepsii (0.8%). Hexyl oleate and octyl oleate were detected for the first time in defensive millipede fluids. The chemical composition of the defensive secretions supports the chemotaxonomic position of the family Blaniulidae in the 'quinone' millipede clade.

Tetradenia riparia (Hochstetter) Codd belongs to the Lamiaceae family and it was introduced in Brazil as an exotic ornamental plant. A previous study showed its antimicrobial, acaricidal and analgesic activities. Two compounds were isolated from essential oil of T. riparia leaves and identified as 9 beta, 13 beta-epoxy-7-abietene (1), a new one, and 6,7-dehydroroyleanone (2), already reported for another plant. The structure of these compounds was determined by spectroscopic analysis and by comparison with literature data. The cytotoxic activities of the essential oil and compounds 1 and 2 were determined by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay, and by tumor cells MDA-MB-435 (human breast carcinoma), HCT-8 (human colon), SF-295 (human nervous system) and HL-60 (human promyelocytic leukemia). The essential oil and compound 1 showed high cytotoxic potential of the cell lines SF-295 (78.06% and 94.80%, respectively), HCT-8 (85.00% and 86.54%, respectively) and MDA-MB-435 (59.48% and 45.43%, respectively). Compound 2 had no cytotoxic activity. The antioxidant activity was determined by 2,2-diphenyl-1-picryl-hydrazyl (DPPH), beta-carotene-linoleic acid system and 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays. The inhibitory concentration (IC50 in mu g mL(-1)) for essential oil and compound 2 was, respectively 15.63 and 0.01 for DPPH; 130.1 and 109.6 for beta-carotene-linoleic acid and 1524 and 1024 for ABTS. Compound 1 had no antioxidant activity. By fractioning the oil, it was possible to identify two unpublished compounds: 1 with high cytotoxic potential and 2 with high antioxidant potential.

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.

The examination of the aerial parts, roots, and seeds of the endemic plant Rindera umbellata is reported in this paper for the first time. Phytochemical investigation of R. umbellata led to the isolation and characterization of ten pyrrolizidine alkaloids and eleven fatty acids in the form of triglycerides. Pyrrolizidine alkaloids 1-9 were found in the aerial parts, 7 and 8 in the roots, and 6-10, together with eleven fatty acids, in the seeds of this plant species. The structures of compounds 1-10 were established based on spectroscopic studies (H-1- and C-13-NMR, 2D NMR, IR and CI-MS). After trans-esterification, methyl esters of the fatty acids were analyzed using GC-MS. The effect of lindelofine-N-oxide (7) on tubulin polymerization was determined.