Boban Anđelković

Chemical composition, cytotoxic and antioxidative activities of ethanolic extracts of propolis on HCT-116 cell line.

BACKGROUND Propolis is a complex resinous sticky substance that honeybees collect from buds and exudates of various plants. Owing to its versatile biological and pharmacological activities, propolis is widely used in medicines, cosmetics and foods. The aim of this study was to evaluate the cytotoxic and antioxidative effects of various ethanolic extracts of propolis (EEPs) on human colon cancer cell line HCT-116 and compare them with their composition determined by HPLC-DAD. RESULTS The most abundant flavonoids in all samples were chrysin, pinocembrin and galangin (12.697-40.811 µg mg⁻¹), while the main phenolic acids were caffeic acid, ferulic acid and isoferulic acid. Dose- and time-dependent inhibition of growth of HCT-116 cells was observed for all propolis samples, with IC₅₀ values ranging from 26.33 to 143.09 µg mL⁻¹. Differences in cytotoxic activity of propolis samples were associated with differences in their composition. All EEP samples reduced both superoxide anion radical and nitrite levels and also had strong DPPH-scavenging activity. CONCLUSION All tested propolis samples had pronounced cytotoxic and antioxidative activities.

Defensive Secretions in Three Ground-Beetle Species (Insecta: Coleoptera: Carabidae)

The adults of three ground-beetle species were induced to discharge defensive secretions into vials. The secretions were obtained by CH2Cl2 extraction. Altogether 11 compounds were identified by GC-MS analysis. Calosoma sycophanta possesses 10 defensive compounds, Carabus ullrichii seven, while Abax parallelepipedus has six compounds. Methacrylic, tiglic and isobutyric acids were present in all samples. The first two organic compounds were predominant in the extracts of Abax parallelepipedus. Methacrylic acid and salicylaldehyde were the major compounds in extracts of Calosoma sycophanta. Methacrylic and angelic acids were the major components in extracts of Carabus ullrichii. Propanoic acid was detected for the first time in the family Carabidae and in all animals. 2-Methyl butyric, angelic and benzoic acids were found for the first time in the subfamily Carabinae. Our finding of butyric acid is its first precise identification in the Carabinae subfamily. 2-Methyl butyric, angelic, crotonic, senecioic and benzoic acids were found for the first time in a European groundbeetle species. The compounds detected in the defensive secretions serve as protection against predators.

Metabolomics study of Populus type propolis

&NA; Herein, we propose rapid and simple spectroscopic methods to determine the chemical composition of propolis derived from various Populus species using a metabolomics approach. In order to correlate variability in Populus type propolis composition with the altitude of its collection, NMR, IR, and UV spectroscopy followed by OPLS was conducted. The botanical origin of propolis was established by comparing propolis spectral data to those of buds of various Populus species. An O2PLS method was utilized to integrate two blocks of data. According to OPLS and O2PLS, the major compounds in propolis samples, collected from temperate continental climate above 500 m, were phenolic glycerides originating from P. tremula buds. Flavonoids were predominant in propolis samples collected below 400 m, originating from P. nigra and P. x euramericana buds. Samples collected at 400–500 m were of mixed origin, with variable amounts of all detected metabolites. Graphical abstract Figure. No caption available. HighlightsAltitude variability in Populus type propolis composition is investigated.NMR, IR, and UV spectroscopy methods in combination with OPLS are applied.Botanical origin of propolis is determined.An O2PLS method is used to integrate spectral data of propolis and Populus buds.

'Does my Diet Affect my Perfume?' Identification and Quantification of Cuticular Compounds in Five Drosophila melanogaster Strains Maintained over 300 Generations on Different Diets.

Cuticular hydrocarbons (CHCs) in Drosophila melanogaster represent the basis of chemical communication being involved in many important biological functions. The aim of this study was to characterize chemical composition and variation of cuticular profiles in five D. melanogaster strains. These strains were reared for approximately 300 generations on five diets: standard cornmeal medium and substrates prepared with apple, banana, tomato, and carrot. Differences in quantity and/or quality in CHCs were assumed as a result of activation of different metabolic pathways involved in food digestion and adaptations to the particular diet type.

Chemical Ecology of Cave-Dwelling Millipedes: Defensive Secretions of the Typhloiulini (Diplopoda, Julida, Julidae)

Cave animals live under highly constant ecological conditions and in permanent darkness, and many evolutionary adaptations of cave-dwellers have been triggered by their specific environment. A similar “cave effect” leading to pronounced chemical interactions under such conditions may be assumed, but the chemoecology of troglobionts is mostly unknown. We investigated the defensive chemistry of a largely cave-dwelling julid group, the controversial tribe “Typhloiulini”, and we included some cave-dwelling and some endogean representatives. While chemical defense in juliform diplopods is known to be highly uniform, and mainly based on methyl- and methoxy-substituted benzoquinones, the defensive secretions of typhloiulines contained ethyl-benzoquinones and related compounds. Interestingly, ethyl-benzoquinones were found in some, but not all cave-dwelling typhloiulines, and some non-cave dwellers also contained these compounds. On the other hand, ethyl-benzoquinones were not detected in troglobiont nor in endogean typhloiuline outgroups. In order to explain the taxonomic pattern of ethyl-benzoquinone occurrence, and to unravel whether a cave-effect triggered ethyl-benzoquinone evolution, we classed the “Typhloiulini” investigated here within a phylogenetic framework of julid taxa, and traced the evolutionary history of ethyl-benzoquinones in typhloiulines in relation to cave-dwelling. The results indicated a cave-independent evolution of ethyl-substituted benzoquinones, indicating the absence of a “cave effect” on the secretions of troglobiont Typhloiulini. Ethyl-benzoquinones probably evolved early in an epi- or endogean ancestor of a clade including several, but not all Typhloiulus (basically comprising a taxonomic entity known as “Typhloiulus sensu stricto”) and Serboiulus. Ethyl-benzoquinones are proposed as novel and valuable chemical characters for julid systematics.

Fourier transform infrared spectroscopy provides an evidence of papain denaturation and aggregation during cold storage

Papain is a cysteine protease with wide substrate specificity and many applications. Despite its widespread applications, cold stability of papain has never been studied. Here, we used differential spectroscopy to monitor thermal denaturation process. Papain was the most stabile from 45 °C to 60 °C with ΔG°321 of 13.9±0.3 kJ/mol and Tm value of 84±1 °C. After cold storage, papain lost parts of its native secondary structures elements which gave an increase of 40% of intermolecular β-sheet content (band maximum detected at frequency of 1621 cm(-1) in Fourier transform infrared (FT-IR) spectrum) indicating the presence of secondary structures necessary for aggregation. The presence of protein aggregates after cold storage was also proven by analytical size exclusion chromatography. After six freeze-thaw cycles around 75% of starting enzyme activity of papain was lost due to cold denaturation and aggregation of unfolded protein. Autoproteolysis of papain did not cause significant loss of the protein activity. Upon the cold storage, papain underwent structural rearrangements and aggregation that correspond to other cold denatured proteins, rather than autoproteolysis which could have the commercial importance for the growing polypeptide based industry.

Comparative study of Balkan Sideritis species from Albania, Bulgaria and Macedonia.

Background: Sideritis scardica and S. raeseri are very popular medicinal plants in Albania, Macedonia, and Bulgaria. Objective: Comparison of the phenolic, flavonoid, and diterpene contents and detailed chemical profiles, as well as data on antioxidant activity, comprising plant material from different locations in the Balkan Peninsula. Materials and Methods: Total phenolic and flavonoid contents for the studied Sideritis samples were determined spectrophotometrically. By HPLC-DAD-ESI-MSn analysis, 48 individual phenolic constituents were completely or tentatively identified. Chemical profiling of Sideritis major diterpenes and their quantification was performed by HPLC-ESI-MSD TOF analysis. A combination of two methods (ORAC and DPPH) was applied for evaluation of the antioxidant capacity of Sideritis extracts. Results: The results obtained demonstrated that Sideritis samples (S. scardica and S. raeseri) collected from Albania, Bulgaria and Macedonia are similar in their chemical profiles (they all contain phenylethanoid glycosides, flavonoid 7-O-diglycosides and acetylated flavonoid 7-Odiglycosides, and diterpenes: main components siderol and sideridiol); and antioxidant activity. Although some differences exist, they are not essential. Conclusion: This fact is a good basis for the formulation of range for parameters for quality control of Sideritis extracts and plant material. The studied cultivated plant material is similar in its chemical composition, profile and antioxidant activity to the wild growing samples.

NMR chemical shift and J coupling parameterization and quantum mechanical reference spectrum simulation for selected nerve agent degradation products in aqueous conditions

The spectral parameters of selected nerve agent degradation products relevant to the Chemical Weapons Convention, namely, ethyl methylphosphonate, isopropyl methylphosphonate, pinacolyl methylphosphonate and methylphosphonic acid, were studied in wide range of pH conditions and selected temperatures. The pH and temperature dependence of chemical shifts and J couplings was parameterized using Henderson–Hasselbalch‐based functions. The obtained parameters allowed calculation of precise chemical shifts and J coupling constants in arbitrary pH conditions and typical measurement temperatures, thus facilitating quantum mechanical simulation of reference spectra in the chosen magnetic field strength for chemical verification. Copyright

Metabolic changes in Euphorbia palusrtis latex after fungal infection.

The variations of metabolic profile of the latex of wild-growing Euphorbia palustris was carried out using multivariate analysis of 1H 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 1H and 2D NMR spectra. Principal component analysis of 1H 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.

Phenolic Compounds from Atriplex littoralis and Their Radiation-Mitigating Activity

From the aerial parts of Atriplex littoralis, three new flavonoid glycosides named atriplexins I-III (1-3), a known flavonoid glycoside, spinacetin 3-O-β-d-glucopyranoside (4), arbutin (5), and 4-hydroxybenzyl-β-d-glucopyranoside (6) were isolated. Their structures were elucidated on the basis of detailed spectroscopic analysis, including 1D and 2D NMR (COSY, NOESY, TOCSY, HSQC, HMBC) and HRESITOF MS data. The compounds were tested for in vitro protective effects on chromosome aberrations in peripheral human lymphocytes using a cytochalasin-B-blocked micronucleus (MN) assay in a concentration range of 0.8-7.4 μM of final culture solution. Chromosomal damage was induced by 2 Gy of γ-radiation on binucleated human lymphocytes, and the effects of the compounds were tested 2 to 19 h after irradiation. The frequency of micronuclei (MNi) was scored in binucleated cells, and the nuclear proliferation index was calculated. The highest prevention of in vitro biochemical and cytogenetic damage of human lymphocytes induced by γ-radiation was exhibited by 3 (reduction of MN frequency by 31%), followed by 4 and 6.