The investigation of bioactive secondary metabolites of the methanol extract of eryngium amethystinum

Eryngium amethystinum L. belonging to the Apiaceae family, is a perennial plant distributed in Southeast Europe. Even though this plant is used in traditional medicine, its phytochemical characterization is still incomplete. In this study composition of bioactive constituents of the methanol extract are reported for the first time. By means of the UPLC-LTQ-Orbitrap-MSn method, altogether sixty-three constituents were characterized: eight hydroxybenzoic acid derivatives (7-13, 32), fifteen cinnamic acid derivatives (14, 17-19, 21, 24-26, 28, 30, 39-42 and 44), four flavonoid aglycones (45, 51, 52, 54), twenty-four flavonoid derivatives (23, 27, 29, 31, 33-38, 43, 46-50, 53, 55-59, 61 and 62), three coumarin derivatives (15, 16 and 22) and nine other compounds (1-6, 20, 60 and 63)

Supplementary data for the article: Stojković, D. L. J.; Bacchi, A.; Capucci, D.; Milenković, M. R.; Čobeljić, B.; Trifunović, S. R.; Andelković, K.; Jevtić, V. V.; Vuković, N.; Vukić, M.; et al. Synthesis and Characterization of Palladium(II) Complexes with Glycine Coumarin Derivatives. Journal of the Serbian Chemical Society 2016, 81 (12), 1383–1392. https://doi.org/10.2298/JSC160915087S

Supplementary material for: [https://doi.org/10.2298/JSC160915087S]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2392

Supplementary data for the article: Stojković, D. L. J.; Bacchi, A.; Capucci, D.; Milenković, M. R.; Čobeljić, B.; Trifunović, S. R.; Andelković, K.; Jevtić, V. V.; Vuković, N.; Vukić, M.; et al. Synthesis and Characterization of Palladium(II) Complexes with Glycine Coumarin Derivatives. Journal of the Serbian Chemical Society 2016, 81 (12), 1383–1392. https://doi.org/10.2298/JSC160915087S

Supplementary material for: [https://doi.org/10.2298/JSC160915087S]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2392

Synthesis and characterization of palladium(II) complexes with glycine coumarin derivatives

A Pd(II) complex with methyl 2-([1-{2,4-dioxochroman-3-ylidene}ethyl]amino)acetate was synthesized. The structures of both the ligand and its Pd(II) complex were determined by elemental analysis, and IR and NMR spectroscopy. Recrystallization of the Pd(II) complex from DMF/water solution resulted in its hydrolysis and the formation of the dimethylamine (2-[{1-(2,4-dioxochroman-3-ylidene) ethyl} amino] acetato) palladium(II) complex, the structure of which was determined by elemental analysis, IR, H-1- and C-13-NMR spectroscopy and X-ray analysis.Supplementary material: [http://cherry.chem.bg.ac.rs/handle/123456789/3626

Rare-earth (Gd3+,Yb3+/Tm3+, Eu3+) co-doped hydroxyapatite as magnetic, up-conversion and down-conversion materials for multimodal imaging

Taking advantage of the flexibility of the apatite structure, nano- and micro-particles of hydroxyapatite (HAp) were doped with different combinations of rare earth ions (RE3+ = Gd, Eu, Yb, Tm) to achieve a synergy among their magnetic and optical properties and to enable their application in preventive medicine, particularly diagnostics based on multimodal imaging. All powders were synthesized through hydrothermal processing at T ≤ 200 °C. An X-ray powder diffraction analysis showed that all powders crystallized in P63/m space group of the hexagonal crystal structure. The refined unit-cell parameters reflected a decrease in the unit cell volume as a result of the partial substitution of Ca2+ with smaller RE3+ ions at both cation positions. The FTIR analysis additionally suggested that a synergy may exist solely in the triply doped system, where the lattice symmetry and vibration modes become more coherent than in the singly or doubly doped systems. HAp:RE3+ optical characterization revealed a change in the energy band gap and the appearance of a weak blue luminescence (λex = 370 nm) due to an increased concentration of defects. The "up"- and the "down"-conversion spectra of HAp:Gd/Yb/Tm and HAp:Gd/Eu powders showed characteristic transitions of Tm3+ and Eu3+, respectively. Furthermore, in contrast to diamagnetic HAp, all HAp:RE3+ powders exhibited paramagnetic behavior. Cell viability tests of HAp:Gd/Yb/Tm and HAp:Gd/Eu powders in human dental pulp stem cell cultures indicated their good biocompatibility

Microbiological Quality of Deer Meat Treated with Essential Oil Litsea cubeba

The present study aimed to evaluate deer meat microbiological quality when treated with essential oil (EO) from Litsea cubeba (dissolved in rapeseed oil at concentrations 0.5 and 1%), in combination with vacuum packaging during 20 days of storage of meat at 4 °C. Total viable counts (TVC), coliforms bacteria (CB), lactic acid bacteria (LAB) and Pseudomonas spp. were analysed at day 0, 1, 5, 10, 15 and 20. MALDI-TOF MS Biotyper technology was applied to identify microorganisms isolated from meat. The highest number of TVC at the end of the experiment was 5.50 log CFU/g in the aerobically packaged control group and the lowest number of TVC was 5.17 log CFU/g in the samples treated with 1.0% Litsea cubeba EO. CB were not detected in the samples treated with 1.0% Litsea cubeba EO during the entire storage period. Bacteria of the genus Pseudomonas were detected only in the aerobically and vacuum packaged control group. The highest number of LAB was 2.06 log CFU/g in the aerobic control group, and the lowest number of LAB was 2.01 log CFU/g in the samples treated with 1.0% Litsea cubeba EO on day 20. The most frequently isolated bacteria from deer meat were Pseudomonas ludensis, Pseudomonas corrugata, Pseudomonas fragi, Bacillus cereus, Staphylococcus epidermidis and Sphingomonas leidyi

Matična mliječ i trans-10-hidroksi-2-decenska kiselina inhibiraju migraciju i invaziju stanica kolorektalnog karcinoma

Research background. Acquisition of migratory potential is pivotal for cancer cells, enabling invasion and metastasis of colorectal carcinoma. Royal jelly and its bioactive component trans-10-hydroxy-2-decenoic acid (10H2DA) showed remarkable antimetastatic potential, but the molecular mechanism underlying this activity is unclear. Experimental approach. Identification and quantification of 10H2DA in royal jelly originating from Serbia was done by HPLC method. Cytotoxicity of 10H2DA was measured by tetrazolium dye MTT test in concentration range 1-500 μg/mL after 24 and 72 h. Its effect on the collective and single-cell migration was measured by wound healing and transwell migration assays. Invasive potential of cancer cells was evaluated by a transwell method modified with collagen. Immunofluorescence was used for migratory and invasive protein expression, while the gene expression of these markers was evaluated by quantitative real time polymerase chain reaction (qRT-PCR). All assays were applied on human colorectal carcinoma HCT-116 and SW-480 cell lines and, except for MTT, evaluated after 24 h of treatment with two selected concentrations of royal jelly and 10H2DA. Results and conclusions. According to HPLC, the mass fraction of 10H2DA in royal jelly was 0.92% (m/m). Treatment with 10H2DA showed no cytotoxic effect; however, significant inhibitory potential of royal jelly and 10H2DA on the motility and invasiveness of colorectal cancer cells was observed. More pronounced effect was exerted by 10H2DA, which significantly suppressed collective cell migration and invasiveness of SW-480 cells, as well as single- and collective cell migration and invasive potential of HCT-116 cell line. Treatments increased epithelial markers E-cadherin and cytoplasmic β-catenin in HCT-116 cells, thus stabilizing intercellular connections. In SW-480 cells, 10H2DA increased E-cadherin on protein and gene level, and suppressed epithelial-mesenchymal transition (EMT) markers. In both cell lines, treatments induced significant suppression of promigratory/proinvasive markers: N-cadherin, vimentin and Snail on protein and gene level, which explains decreased migratory and invasive potential of HCT-116 and SW-480 cells. Novelty and scientific contribution. Our study presents new findings and elucidation of royal jelly and 10H2DA molecular mechanism that underlies their antimigratory/antiinvasive activity on colorectal cancer cells. These findings are shown for the first time indicating that these natural products are a valuable source of anticancer potential and should be reconsidered for further antitumour therapy.Pozadina istraživanja. Sposobnost migracije stanica jе ključna za invaziju i mеtastaziranje kolorektalnog karcinoma. Matična mlijеč te njezin bioaktivni sastojak trans-10-hidroksi-2-dеcеnska kisеlina (10H2DA) imaju izuzеtan antimеtastatski potеncijal, no molеkularni mеhanizam ovе aktivnosti još uvijek nije jasan. Eksperimentalni pristup. Prisutnost i količina 10H2DA u matičnoj mlijеči porijеklom iz Srbijе utvrđeni su mеtodom HPLC. Citotoksičnost 10H2DA ispitana jе nakon 24 i 72 h pomoću MTT tеsta s 1−500 μg/mL tetrazolijeve soli. Utjecaj 10H2DA na kolеktivnu migraciju i onu pojedinih stanica određen je praćenjem procesa cijeljenja rana te migracije stanica na Transwell pločama. Invazivni potеncijal stanica karcinoma ispitan jе na Transwell pločama s kolagеnom. Za određivanje еksprеsije protеina uključenih u procese migracije i invazije upotrijebljena jе imunofluorеscеntna metoda, dok jе gеnska еksprеsija tih markеra procijenjena kvantitativnom lančanom rеakcijom polimеrazе u stvarnom vrеmеnu (qRT-PCR). U svim su testovima korištene dvije stanične linije humanog kolorеktalnog karcinoma: HCT-116 i SW-480, tretirane s dvijе odabranе koncеntracijе matične mlijеči i 10H2DA, čiji je učinak mjeren 24 h nakon tretmana, osim u MTT testu. Rezultati i zaključci. Metodom HPLC utvrđeno je da matična mliječ sadržava 0,92 % (m/m) 10H2DA. Ispitivanjem utjecaja 10H2DA na stanice karcinoma utvrđeno je da kiselina nijе imala citotoksični učinak, no opažen jе znatan potеncijal matične mlijеči i 10H2DA da inhibiraju pokrеtljivost i invazivnost stanica karcinoma dеbеlog crijеva. Izražеnije je djelovanje imala 10H2DA, koja jе bitno smanjila kolеktivnu migraciju i invazivnost SW-480 stanica, kao i kolеktivnu migraciju, migraciju pojеdinih stanica i invazivni potеncijal stanične linijе HCT-116. Nakon obrade povеćala se ekspresija еpitеlnih markеra E-kadhеrina i citoplazmatskog β-katеnina u HCT-116 stanicama, što je dovelo do stabilizacije međustaničnih vеza. U stanicama SW-480 je 10H2DA povеćala ekspresiju E-kadhеrina na razini proteina i gena te inhibirala markеrе еpitеlno-mеzеnhimalnе tranzicijе (EMT). Obrada matičnom mliječi i 10H2DA je u objе stanične linijе u većoj mjeri potaknula suprеsiju promigracijskih/proinvazivnih markеra N-kadhеrina, vimеntina i Snail-a na razini gena i proteina, što objašnjava smanjеn migracijski i invazivni potеncijal HCT-116 i SW-480 stanica nakon obrade. Novina i znanstveni doprinos. Ovo istraživanje donosi nove spoznaje i pojašnjava molеkularni mеhanizam antimigracijskog/antiinvazivnog djelovanja matične mlijеči i 10H2DA na stanice raka dеbеlog crijеva. Rezultati po prvi put pokazuju da su ovi prirodni proizvodi vrijеdan izvor s antikancеrogеnim potеncijalom i da ih trеba pomnije razmotriti za buduću antitumorsku tеrapiju

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 mu gmg(-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 mu gmL(-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.Peer-reviewed manuscript: [http://cherry.chem.bg.ac.rs/handle/123456789/3472

Chemical Composition of Various <i>Nepeta cataria</i> Plant Organs’ Methanol Extracts Associated with <i>In Vivo</i> Hepatoprotective and Antigenotoxic Features as well as Molecular Modeling Investigations

This report summarizes the chemical composition analysis of Nepeta cataria L. flower, leaf, and stem methanol extracts (FME, LME, SME, respectively) as well as their hepatoprotective and antigenotoxic features in vivo and in silico. Herein, Wistar rat liver intoxication with CCl4 resulted in the generation of trichloromethyl and trichloromethylperoxy radicals, causing lipid peroxidation within the hepatocyte membranes (viz. hepatotoxicity), as well as the subsequent formation of aberrant rDNA adducts and consequent double-strand break (namely genotoxicity). Examined FME, LME, and SME administered orally to Wistar rats before the injection of CCl4 exerted the most notable pharmacological properties in the concentrations of 200, 100, and 50 mg/kg of body weight, respectively. Thus, the extracts’ hepatoprotective features were determined by monitoring the catalytic activities of enzymes and the concentrations of reactive oxidative species, modulating the liver redox status. Furthermore, the necrosis of hepatocytes was assessed by means of catalytic activities of liver toxicity markers. The extracts’ antigenotoxic features were quantified using the comet assay. Distinct pharmacological property features may be attributed to quercitrin (8406.31 μg/g), chlorogenic acid (1647.32 μg/g), and quinic acid (536.11 μg/g), found within the FME, rosmarinic acid (1056.14 μg/g), and chlorogenic acid (648.52 μg/g), occurring within the LME, and chlorogenic acid (1408.43 μg/g), the most abundant in SME. Hence, the plant’s secondary metabolites were individually administered similar to extracts, upon which their pharmacology in vivo was elucidated in silico by means of the structure-based studies within rat catalase, as a redox marker, and rat topoisomerase IIα, an enzyme catalyzing the rat DNA double-strand break. Conclusively, the examined N. cataria extracts in specified concentrations could be used in clinical therapy for the prevention of toxin-induced liver diseases

<i>Thymus zygis,</i> Valuable Antimicrobial (In Vitro and In Situ) and Antibiofilm Agent with Potential Antiproliferative Effects

With the growing issues of food spoilage, microbial resistance, and high mortality caused by cancer, the aim of this study was to evaluate T. zygis essential oil (TZEO) as a potential solution for these challenges. Here, we first performed GC/MS analysis which showed that the tested TZEO belongs to the linalool chemotype since the abundance of linalool was found to be 38.0%. Antioxidant activity assays showed the superiority of TZEO in neutralizing the ABTS radical cation compared to the DPPH radical. The TZEO was able to neutralize 50% of ABTS•+ at the concentration of 53.03 ± 1.34 μg/mL. Antimicrobial assessment performed by employing disc diffusion and minimal inhibitory concentration assays revealed TZEO as a potent antimicrobial agent with the highest inhibition activity towards tested gram-negative strains. The most sensitive on the treatment with TZEO was Enterobacter aerogenes showing an MIC 50 value of 0.147 ± 0.006 mg/mL and a MIC 90 value of 0.158 ± 0.024 mg/mL. Additionally, an in situ analysis showed great effects of TZEO in inhibiting gram-negative E. coli, P. putida, and E. aerogenes growing on bananas and cucumbers. Treatment with the TZEO vapor phase in the concentration of 500 μg/mL was able to reduce the growth of these bacteria on the food models to the extent > 90%, except for E. coli growth on the cucumber, which was reduced to the extent of 83.87 ± 4.76%. Furthermore, a test on the antibiofilm activity of the tested essential oil revealed its biofilm prevention effects against Salmonella enterica which forms biofilms on plastic and stainless-steel surfaces. Performed tests on the TZEO effects towards cell viability showed no effects on the normal MRC-5 cell line. However, the results of MTT assay of TZEO effects on three cancer cell lines (MDA-MB-231, HCT-116, and K562) suggest that TZEO exerted the strongest effects on the inhibition of the viability of MDA-MB-231 cells, especially after long-term treatment in the highest concentration applied with reducing the viability of the cells to 57%. Additionally, results of NBT and Griess assays suggest that TZEO could be a convenient candidate for future testing for developing novel antitumor therapies