10 research outputs found
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)
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
Shikonin derivatives from onsoma visianii decrease expression of phosphorylated stat3 in leukemia cells and exert antitumor activity
Antitumor effects of shikonins on chronic lymphocytic leukemia (CLL) and B-cell pro-lymphocytic leukemia (B-PLL) are mostly unexplored. The antitumor activity of shikonins, isolated from Onosma visianii Clem (Boraginaceae), in BCL1, mouse CLL cells and JVM-13, human B-PLL cells was explored in this study. The cytotoxicity of shikonin derivatives was measured by an MTT test. Cell death, proliferation, cell cycle, and expression of molecules that control these processes were analyzed by flow cytometry. Expression of STAT3-regulated genes was analyzed by real-time q-RT-PCR (Quantitative Real-Time Polymerase Chain Reaction). The antitumor effects of shikonin derivatives in vivo were analyzed, using flow cytometry, by detection of leukemia cells in the peripheral blood and spleens of mice intravenously injected with BCL1 cells. The two most potent derivatives, isobutyrylshikonin (IBS) and α-methylbutyrylshikonin (MBS), induced cell cycle dis-turbances and apoptosis, inhibited proliferation, and decreased expression of phospho-STAT3 and downstream-regulated molecules in BCL1 and JVM-13 cells. IBS and MBS decreased the percentage of leukemia cells in vivo. The link between the decrease in phosphorylated STAT3 by MBS and IBS and BCL1 cell death was confirmed by detection of enhanced cell death after addition of AG490, an inhibitor of Jak2 kinase. It seems that IBS and MBS, by decreasing STAT3 phosphorylation, trigger apoptosis, inhibit cell proliferation, and attenuate leukemia cell stemness
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
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
Chemical composition, in vitro and in situ antimicrobial and antibiofilm activities of syzygium aromaticum (Clove) essential oil
The essential oil of Syzygium (S.) aromaticum (CEO) is known for its good biological activity. The aim of the research was to evaluate in vitro and in situ antimicrobial and antibiofilm activity of the essential oil produced in Slovakia. The main components of CEO were eugenol 82.4% and (E)-caryophyllene 14.0%. The antimicrobial activity was either weak or very strong with inhibition zones ranging from 4.67 to 15.78 mm in gram-positive and gram-negative bacteria and from 8.22 to 18.56 mm in yeasts and fungi. Among the tested bacteria and fungi, the lowest values of MIC were determined for Staphylococcus (S.) aureus and Penicillium (P.) expansum, respectively. The vapor phase of CEO inhibited the growth of the microscopic filamentous fungi of the genus Penicillium when tested in situ on bread. The strongest effect of mycelia inhibition in a bread model was observed against P. expansum at concentrations of 250 and 500 µL/mL. The best antimicrobial activity of CEO in the carrot model was found against P. chrysosenum. Differences between the mass spectra of Bacillus (B.) subtilis biofilms on the tested surfaces (wood, glass) and the control sample were noted from the seventh day of culture. There were some changes in mass spectra of Stenotrophomonas (S.) maltophilia, which were observed in both experimental groups from the fifth day of culture. These findings confirmed the impact of CEO on the protein structure of older biofilms. The findings indicate that, besides being safe and sensorially attractive, S. aromaticum has antimicrobial activity, which makes it a potential substitute for chemical food preservatives
<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
Synthesis and Biological Screening of New 4-Hydroxycoumarin Derivatives and Their Palladium(II) Complexes
Two newly synthesized 4-hydroxycoumarin bidentate ligands (L1 and L2) and their palladium(II) complexes (C1 and C2) were screened for their biological activities, in vitro and in vivo. Structures of new compounds were established based on elemental analysis, 1H NMR, 13C NMR, and IR spectroscopic techniques. The obtained compounds were tested for their antioxidative and cytotoxic activities and results pointed to selective antiradical activity of palladium(II) complexes towards •OH and -•OOH radicals and anti-ABTS (2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) cation radical) activity comparable to that of ascorbate. Results indicated the effect of C1 and C2 on the enzymatic activity of the antioxidative defense system. In vitro cytotoxicity assay performed on different carcinoma cell lines (HCT166, A375, and MIA PaCa-2), and one healthy fibroblast cell line (MRC-5) showed a cytotoxic effect of both C1 and C2, expressed as a decrease in carcinoma cells’ viability, mostly by induction of apoptosis. In vivo toxicity tests performed on zebrafish embryos indicated different effects of C1 and C2, ranging from adverse developmental effect to no toxicity, depending on tested concentration. According to docking studies, both complexes (C1 and C2) showed better inhibitory activity in comparison to other palladium(II) complexes
Anti-Tumor Mechanisms of Novel 3-(4-Substituted Benzyl)-5-Isopropil-5- Phenylhydantoin Derivatives in Human Colon Cancer Cell Line
Brašno - Kruh '13
The "Flour-Bread '13“ Congress topics were the following: breeding and quality of cereal grains, grain storage and milling technology, analytical and rheological methods, baking technology, improvers and additives, starch and modified starch, extrusion and pasta production, biscuit and pastry products, nutritional quality of cereals, cereal food safety and cereal based functional foods