Kemijski sastav i biološka aktivnost eteričnog ulja i ekstrakta sjemenki visokog dragoljuba (Tropaeolum majus L. var. altum)

Research background. Plant Tropaeolum majus L. (garden nasturtium) belongs to the family Tropaeolaceae and contains benzyl glucosinolate. The breakdown product of benzyl glucosinolate, benzyl isothiocyanate (BITC), exhibits various biological activities such as antiproliferative, antibacterial and antiinflammatory. In order to optimize the content of biologically active volatile compounds in plant extract and essential oil, the use of appropriate extraction technique has a crucial role. Experimental approach. The current study investigates the effect of two modern extraction methods, microwave-assisted distillation (MAD) and microwave hydrodiffusion and gravity (MHG), on the chemical composition of volatile components present in the essential oil and extract of garden nasturtium (T. majus L. var. altum) seeds. Investigation of the biological activity of samples (essential oil, extract and pure compounds) was focused on the antiproliferative effect against different cancer cell lines: cervical cancer cell line (HeLa), human colon cancer cell line (HCT116) and human osteosarcoma cell line (U2OS), and the antibacterial activity which was evaluated against the growth and adhesion of Staphylococcus aureus and Escherichia coli to polystyrene surface. Results and conclusions. Essential oil and extract of garden nasturtium (T. majus) seeds were isolated by two extraction techniques: MAD and MHG. BITC and benzyl cyanide (BCN) present in the extract were identified by gas chromatography-mass spectrometry. Essential oil of T. majus showed higher antiproliferative activity (IC50<5 µg/mL) than T. majus extract (IC50<27 µg/mL) against three cancer cell lines: HeLa, HCT116 and U2OS. BITC showed much higher inhibitory effect on all tested cells than BCN. The essential oil and extract of T. majus showed strong antimicrobial activity against S. aureus and E. coli. Novelty and scientific contribution. This work represents the first comparative report on the antiproliferative activity of the essential oil and extract of T. majus seeds, BITC and BCN against HeLa, HCT116 and U2OS cells as well as their antimicrobial activity against S. aureus and E. coli. This study demonstrates that the essential oil of T. majus seeds exhibits stronger antiproliferative and antimicrobial activity than the plant extract.Pozadina istraživanja. Biljka dragoljub (Tropaeolum majus L.) pripada porodici Tropaeolaceae i sadržava benzil-glukozinolat. Razgradni produkt benzil-glukozinolata, benzil-izotiocijanat (BITC), ispoljava različite biološke aktivnosti, poput antiproliferativne, antibakterijske i protuupalne. U optimiranju sadržaja biološki aktivnih, hlapljivih spojeva u biljnom ekstraktu i eteričnom ulju ključnu ulogu ima primjena odgovarajuće tehnike ekstrakcije. Eksperimentalni pristup. U ovom je istraživanju ispitan učinak dviju naprednih tehnika ekstrakcije, destilacije potpomognute mikrovalovima (MAD) i mikrovalne hidrodifuzije i gravitacije (MHG), na kemijski sastav hlapljivih komponenti prisutnih u eteričnom ulju i ekstraktu sjemenki visokog dragoljuba (T. majus L. var. altum). Istraživanje biološke aktivnosti uzoraka (eteričnog ulja, ekstrakta i čistih spojeva) bilo je usmjereno na njihov antiproliferativni učinak na stanične linije raka grlića maternice (HeLa), raka debelog crijeva (HCT116) i osteosarkoma (U2OS), te na njihov učinak na suzbijanje rasta i adhezije bakterija Staphylococcus aureus i Escherichia coli na polistirenskoj površini. Rezultati i zaključci. Eterično ulje i ekstrakt sjemenki dragoljuba (T. majus) izolirani su dvjema tehnikama ekstrakcije, MAD i MHG. Prisutnost benzil-izotiocijanata i benzil-cijanida utvrđena je plinskom kromatografijom spregnutom s masenom spektrometrijom. Eterično ulje dragoljuba imalo je izraženiji antiproliferativni učinak (IC50<5 µg/mL) na stanične linije raka HeLa, HCT116 i U2OS od ekstrakta biljke (IC50<27 µg/mL). Benzil-izotiocijanat je imao mnogo veći inhibitorni učinak na sve ispitane stanice raka od benzil-cijanida. Eterično ulje i ekstrakt dragoljuba imali su snažan antimikrobni učinak na S. aureus i E. coli. Novost i znanstveni doprinos. Ovaj rad predstavlja prvo komparativno izvješće o antiproliferativnom djelovanju eteričnog ulja i ekstrakta sjemenki dragoljuba, te čistih spojeva BITC i BCN na HeLa, HCT116 i U2OS stanice, kao i njihovom antimikrobnom učinku na S. aureus i E. coli. Rezultati istraživanja pokazuju da eterično ulje sjemenki dragoljuba ima jače antiproliferativno i antimikrobno djelovanje od ekstrakta te biljke

Genome structure, chemical composition and biological activity of triploid onion Allium × cornutum Clementi ex Visiani

Allium × cornutum Clementi ex Visiani jedinstvena je forma luka, koja se pod nazivom 'Ljutika' tradicionalno uzgaja i koristi u ljudskoj prehrani u obalnoj i otočnoj Hrvatskoj. Otporna je na sušu i hladnoću, te prilagođena oskudnom, kamenitom tlu. Ovim radom željela se detaljnije istražiti struktura genoma te vrste, rasvijetliti nutritivni sastav te provjeriti njezinu biološku aktivnost. Provedene molekularno-filogenetske analize ribosomskih gena, 18S-5.8S-26S rDNA i 5S rDNA, otkrile su da je A. × cornutum (2n = 3x = 24) jedinstveni hibrid triju vrsta lukova: crvenog luka, Allium cepa L. i dva azijska luka, Allium pskemense B. Fedtsch i Allium roylei Stearn. Po jedan haploidni genom (x = 8) svake od triju roditeljskih vrsta identificiran je i vizualiziran na mitotskim i mejotičkim kromosomima A. × cornutum metodom fluorescentne hibridizacije in situ. Analize kloroplastnih i mitohondrijskih gena, pokazuju da A. × cornutum posjeduje jedinstveni tip sterilne citoplazme CMS-S, važne u oplemenjivanju lukova. Kemijske analize metanolnih ekstrakta A. × cornutum i A. cepa izvršene metodom HPLC-a, ukazuju na povećan sadržaj flavonoida (flavonola i antocijanina) u A. × cornutum, što može biti objašnjenje i nešto boljeg bioaktivnog profila triploida u odnosu na A. cepa. Obje vrste pokazuju značajnu antioksidativnu, antigenotoksičnu i antiproliferacijsku aktivnost, što ih potvrđuje kao važan izvor korisnih fitonutrijenata koji mogu doprinijeti zaštiti i očuvanju ljudskog zdravlja.Allium × cornutum Clementi ex Visiani is unique onion type, traditionally cultivated in South and coastal Croatia under the name 'Ljutika' and used in human diet. It is resistant to cold weather conditiones and drought, and adapted to growth on poor stony ground. This paper attempts to explore genome structure, shed light on the nutritional composition and check biological activity of this species. Molecular-cytogenetic and phylogenetic analysis of two ribosomal genes, 18S-5.8S-26S rDNA and 5S rDNA revealed that 'Ljutika' is a unique hybrid of three onion species: common onion, Allium cepa L., and two Asian onions, Allium pskemense B. Fedtsch and Allium roylei Stearn. One haploid genome (x = 8) of each of the three parent species was identified and visualized on mitotic and meiotic chromosomes of A. × cornutum by fluorescent in situ hybridization. Chloroplast and mitochondrial gene analysis showed that 'Ljutika' possesses a sterile CMS-S type of cytoplasm, important for onion breeding. Phytochemical analysis determined by HPLC showed that methanolic extracts of A. × cornutum and A. cepa showed slightly higher content of flavonoids (flavonols and anthocyanins) in A. × cornutum, which may be cause of its higher bioactive profile in compare to A. cepa. Both species showed a significant antioxidant, antigenotoxic and antiproliferative activity which makes them an important sources of phytonutrients that can contribute to the protection and preservation of human health

Comparison of Organosulfur and Amino Acid Composition between Triploid Onion Allium cornutum Clementi ex Visiani, 1842, and Common Onion Allium cepa L., and Evidences for Antiproliferative Activity of Their Extracts

Species that belong to the genus Allium have been widely used for human food and traditional medicine. Their beneficial health effects, as well as the specific aroma, are associated with their bioactive chemical compounds, such as sulfur compounds and flavonoids. Gas chromatography and mass spectrometry (GC&ndash;MS) and reverse-phase high-performance liquid chromatography (reverse-phase HPLC) were used to identify organosulfur and amino acid content of triploid hybrid onion, Allium cornutum Clement ex Visiani, 1842, and common onion, Allium cepa L. Allium extracts were tested for their antiproliferative activity in three human cancer cell lines (HeLa, HCT116, and U2OS). DNA fragmentation and DAPI staining analysis were performed on HeLa cells to evaluate the effect of extracts on DNA damage and cell morphology. The mRNA expression of p53, Bax, and Caspase-3 genes involved in apoptosis were analyzed by real-time PCR. Using GC&ndash;MS, 27 compounds were found in two Allium species headspaces. Differences were noted among the main compound abundance in the headspace (although the major thiols and disulfides were qualitatively identic in both Allium species) and dipropyl disulfide, diisopropyl trisulfide, and (Z)-prop-1-enyl propyl trisulfide were predominant sulfides. Identification of amino acids and their quantities were determined by reverse-phase HPLC. Most abundant amino acids in both onions were arginine (Arg) and glutamic acid (Glu). The results of cytotoxicity testing confirmed antiproliferative effects of both species. The DNA fragmentation assay, DAPI staining and real time PCR analysis confirmed that A. cornutum and A. cepa extracts induced apoptosis in HeLa cells. This study presents the evidence for possible therapeutic use of A. cornutum and A. cepa extracts against human cervical carcinoma cell line

Chemical Composition and New Biological Activities of Essential Oil and Hydrosol of Hypericum perforatum L. ssp. veronense (Schrank) H. Lindb

The chemical profile, antiproliferative, antioxidant and antiphytoviral activities of the species Hypericum perforatum ssp. veronense (Schrank) H. Lindb. (Clusiaceae) were investigated. Free volatiles were isolated and the chemical composition was determined in the lipophilic fraction (essential oil) and for the first time in the water fraction (hydrosol). The aim is to provide phytochemical data for H. perforatum ssp. veronense useful for distinguishing ssp. veronense from ssp. angustifolium, as there are taxonomic disagreements between them and the composition of the secretory products may be helpful in this respect. In the essential oil, the most abundant compounds identified were α-pinene and n-nonane, while in the hydrosol, myrtenol, carvacrol and α-pinene were the most abundant. Overall, the class of monoterpenes and oxygenated monoterpenes dominated in the EO and hydrosol samples. The essential oil showed high antioxidant activity, in contrast to the antiproliferative activity, where the hydrosol showed exceptional activity against three cancer cell lines: Hela (cervical cancer cell line), HCT116 (human colon cancer cell line) and U2OS (human osteosarcoma cell line). Both the essential oil and hydrosol showed antiphytoviral activity against tobacco mosaic virus infection on the local host plants. This is the first report dealing with biological activities of hydrosol of H. perforatum ssp. veronense, and the obtained results suggest that this traditional medicinal plant is a valuable source of volatiles with promising antiproliferative, antioxidant and antiphytoviral activities

Chemical Composition and Biological Activity of Allium cepa L. and Allium × cornutum (Clementi ex Visiani 1842) Methanolic Extracts

Here, we report a comparative study of the phytochemical profile and the biological activity of two onion extracts, namely Allium cepa L. and Allium × cornutum (Clementi ex Visiani 1842), members of the family Amaryllidaceae. The identification of flavonoids and anthocyanins, and their individual quantities, was determined by high-performance liquid chromatography (HPLC). The potency of both extracts to scavenge free radicals was determined by the DPPH (2,2′-diphenyl-1-picrylhydrazyl) radical-scavenging activity and oxygen radical absorbance capacity (ORAC) methods. The DNA protective role was further tested by the single-cell gel electrophoresis (COMET) assay and by Fenton’s reagent causing double-strand breaks on the closed circular high copy pUC19 plasmid isolated from Escherichia coli. In the presence of both extracts, a significant decrease in DNA damage was observed, which indicates a protective role of Allium cepa and Allium × cornutum on DNA strand breaks. Additionally, cytotoxicity was tested on glioblastoma and breast cancer cell lines. The results showed that both extracts had antiproliferative effects, but the most prominent decrease in cellular growth was observed in glioblastoma cells

Free Volatile Compounds of Veronica austriaca ssp. jacquinii (Baumg.) Eb. Fisch. and Their Biological Activity

The composition of free volatile compounds of essential oils (EO) and hydrosols (Hy) from four different localities of the species Veronica austriaca ssp. jacquinii (Baumg.) Eb. Fisch. were analyzed by gas chromatography coupled with mass spectrometry. In the EOs, the most abundant compounds identified were hexahydrofarnesyl acetone (23.34–52.56%), hexadecanoic acid (palmitic acid, 26.71–58.91%) and octadecanol acetate (0–6.24%). The hydrosols were characterized by high abundance of methyl eugenol (23.35–57.93%), trans-p-mentha-1(7),8-dien-2-ol (5.24–7.69%) and thymol (3.48–9.45%). Glandular trichomes were analyzed using SEM (Scanning Electron Microscopy), as they are the sites of synthesis of free volatile compounds. We have detected glandular trichomes, consisting of a one stalk cell and two elliptically shaped head cells, and non-glandular (unbranched, bi-cellular to multicellular) trichomes on stems, leaves and the sepals. Data for volatile compounds from EOs and hydrosols were analyzed using Principal Component Analyses (PCA) to demonstrate variations in the composition of the volatile compounds identified. Isolated samples of EO and hydrosols were analyzed for their antioxidant activity using two methods, DPPH (2,2-diphenyl-1-picrylhydrazyl) and ORAC (Oxygen Radical Absorbance Capacity). The essential oils showed higher antioxidant activity than the hydrosols in ORAC method, but lower activity by the DPPH method. The isolates were also tested for their antiproliferative activity on different types of cancer cells and also on two lines of healthy cells, and the results showed that the extracts were not toxic to the cell lines tested. Total polyphenols, total tannins, total flavonoids and total phenolic acids were also analyzed and determined spectrophotometrically. The free volatile compounds of Veronica austriaca ssp. jacquinii can be considered as a safe natural product

Picoplankton diversity in an oligotrophic and high salinity environment in the central Adriatic Sea

Abstract By combining qualitative 16S metabarcoding and quantitative CARD-FISH methods with neural gas analysis, different patterns of the picoplankton community were revealed at finer taxonomic levels in response to changing environmental conditions in the Adriatic Sea. We present the results of a one-year study carried out in an oligotrophic environment where increased salinity was recently observed. We have shown that the initial state of community structure changes according to environmental conditions and is expressed as qualitative and quantitative changes. A general pattern of increasing diversity under harsh environmental conditions, particularly under the influence of increasing salinity at the expense of community abundance was observed. Considering the trend of changing seawater characteristics due to climate change, this study helps in understanding a possible structural change in the microbial community of the Adriatic Sea that could affect higher levels of the marine food web