le antocianine sono una classe di metaboliti secondari nelle piante, appartenenti ai flavonoidi, categoria di molecole antiossidanti utili per la salute di piante, animali e umana. Sono ampiamente studiati nelle Leguminosae poiché implicati in numerose funzioni fisiologiche. In questa tesi, due wild-types e cinque mutanti di Medicago truncatula, coinvolti nel metabolismo dei flavonoidi, sono stati caratterizzati a livello fenotipico, fisiologico e molecolare. La principale differenza tra i wild-type e i mutanti è la riduzione del contenuto di antocianine nelle foglie. In seguito all'esposizione alle radiazioni UV-B, è stata analizzata la loro risposta, evidenziando un diverso comportamento. Inoltre, il metabolismo delle antocianine è stato studiato anche a livello di regolazione genica e un nuovo gene myb è stato isolato e caratterizzato, risultando più espresso nei tessuti che accumulano antocianine. / Anthocyanins are secondary metabolites in plants. They belong to a class of flavonoids synthesized via the phenylpropanoid pathway and they are antioxidant molecules important for plant, animal and human health. Flavonoids are widely studied in legume because they are implicated in several biological and physiological functions. In this thesis seven genotypes (two wild-types and five indipendent mutants) of Medicago truncatula (the model legume plant) affected in flavonoid metabolism have been characterized at phenotypic, physiological and molecular level. The main difference between wild-types and mutants, is the reduction in anthocyanin content in leaves. The anthocyanin accumulation during the leaf life, the flowering time and the fruit formation have been registered and compared. The two wild-types contain 6 μg cyanidin.mg-1 of fresh weight, while in the mutants the anthocyanin amount ranged from 0.12 μg cyanidin mg-1 to 2 μg cyanidin mg-1 of fresh weight. After HPLC-DAD-MS/MS analysis, the main anthocyanin present in the two wild-types is the cyanidin-3-O feruloyl. The Expression profile of genes codifying enzymes and transcriptional factor involved in flavonoid biosynthesis has been investigated. RT-PCR and qPCR results show different possible pathways of reduction of anthocyanins in the five mutants. These mutants, have been exposed to UV-B radiations and their response has been investigated measuring the chlorophyll fluorescence parameters (Fv/Fm, qP and NPQ) in untreated plants, during treatment (after 4hrs and 14hrs of treatment) and in the recovering phase. All genotypes, regardless of the anthocyanins amount, showed a decrease of the photosyntetic efficiency after 14hrs of treatment. This indicates a marginal role of these pigments in the oxidative damages protection. Mutants do not response in the same manner to the UV—B exposure and the anthocyanin amount does not increase equally in all genotypes. The anthocyanin metabolism is studied also at the gene regulation level. A novel Myb gene (MtMYBA) involved in anthocyanin pathway has been isolated and characterized. This gene is more expressed in tissues which accumulate anthocyanins in M. truncatula plants. The functional analysis has been investigated overexpressing this Myb gene under the control of 35S promoter. This construct has been used to transform Arabisopsis thaliana and Medicago truncatula plants. In addition, the MtMYBA promoter has been cloned in a plasmids containing GUS and GFP reporter genes.

<p>Bosiljak (Ocimum basilicum L.) pripada familiji Lamiaceae i jedna je od najrasprostranjenijih i najče&scaron;će gajenih biljaka &scaron;irom sveta. Njegova upotreba je &scaron;iroko rasprostranjena zbog bogatog sadržaja i skladnog odnosa vitamina, minerala kao i različitih fenolnih jedinjenja kao glavnih nosilaca antioksidativne aktivnosti. Poslednjih godina velika pažnja je usmerena ka fenolnim fitokomponentama kao potencijalnim promoterima zdravlja. Ciljevi ovog rada bili su ispitivanje kvalitativnih i kvantitativnih karakteristika, kao i biohemijskih aktivnosti ekstrakata bosiljka dobijenih različitim rastvaračima, kao i ispitivanje uticaja farmaceutsko-tehnolo&scaron;ke formulacije u obliku mikrovezikula ekstrakta bosiljka na glikemijski, lipidni i oksido-redukcioni status kod oglednih životinja, u odnosu na vodeni ekstrakt. In vitro ispitivanja su uključivala analizu 100 ekstrakata dobijenih različitim rastvaračima i stepenom usitnjenosti. Ukupni fenolni i flavonoidni sadržaj, kao i antioksidativna aktivnost određena je spektrofotometrijskom metodom. Takođe, kvantifikovane i kvalifikovane su fenolne komponente (fenolne kiseline i flavonoidi) primenom visokoefikasne tečne hromatografije (HPLC). In vivo ispitivanje je rađeno na 84 albino laboratorijska pacova soja Wistar. Per os su primenjivani vodeni ekstrakt bosiljka, rastvor natrijumove soli monoketoholne kiseline, njihove kombinacije, kao i ekstrakt u obliku farmaceutsko-tehnolo&scaron;ke formulacije, mikrovezikula. Posle sedmodnevnog tretmana, merene su koncentracije glukoze u krvi, a nakon žrtvovanja, u serumu su određeni parametri lipidnog statusa i antioksidativnog stresa. Ex vivo analizama procenjivani su efekti ekstrakta bosiljka na enzimske i neenzimske parametre antioksidativnog odbrambenog sistema i oksidativne modifikacije lipida. Praćen je i uticaj tretmana na akutno o&scaron;tećenje jetre usled primene paracetamola. Svi analizirani ekstrakti bosiljka su pokazali prisustvo velikog broja fenolnih jedinjenja iz klase fenolnih kiselina i flavonoida. Detektovane i kvantifikovane aktivne komponente bile su fenolne kiseline: hlorogenska, p-hidroksibenzoeva, kafena, vanilinska, ferulna, rozmarinska i cimetna, kao i apigenin, kvercetin, naringenin i rutin kao predstavnici flavonoidnih jedinjenja. Primena ekstrakta bosiljka i natrijumove soli monoketoholne kiseline, same ili u kombinaciji, a takođe i u farmaceutsko-tehnolo&scaron;koj formulaciji, pokazale su smanjenja o&scaron;tećenja tokom oksidativnog stresa, značajno antidijabetesno delovanje, povoljan uticaj na lipidni status i protektivni efekat na funkcije jetre i bubrega. Na osnovu dobijenih rezultata može se zaključiti da ekstrakt bosiljka sadrži značajnu količinu fenolnih jedinjenja odgovornih za antioksidativnu aktivnost. Vodeni ekstrakt bosiljka, sam ili u kombinaciji sa monoketoholnom kiselinom, i u obliku farmaceutsko-tehnolo&scaron;ke formulacije snižava nivo glukoze u krvi, ubrzava obnovu &beta; ćelija, ima povoljan efekat na lipidni status i ne dovodi do toksičnih promena na jetri i bubrezima kod eksperimentalnih životinja.</p> / <p>Basil (Ocimum basilicum L.) belongs to the family Lamiaceae and represents one of the most widespread and most commonly cultivated plant worldwide. Its use is widespread due to its rich content and harmonious relationship of vitamins, minerals and various phenolic compounds which are responsible for the antioxidant activity. In recent years, great attention is directed to phenolic phytocomponents as potential promoters of health. The objectives of this study were to test the qualitative and quantitative characteristics, as well as the biochemical activity of basil extracts obtained by various solvents and investigate the impact of pharmaceutical technological formulation of basil extract on glycemic, lipid and oxidation-reduction status of the aqueous extract in the experimental animals. In vitro studies have included 100 extracts obtained by various solvents and degrees of fragmentation. Total phenolic and flavonoid content, as well as antioxidant activity was determined by spectrophotometric method. Also, phenolic compounds (phenolic acids and flavonoids) were qualified and quantified using high performance liquid chromatography (HPLC). In vivo testing was performed on 84 laboratory albino Wistar rats. The aqueous extract of basil, a solution of sodium salt of monoketocholic acid, their combinations, as well as an extract in the form of the pharmaceutical technological formulation were administered per os. After seven-day-treatment, the concentrations of glucose were measured in the blood and after sacrificing of animals, the lipid and antioxidative parameters were determined in serum. Ex vivo analysis assessed the effect of the extract of basil on the parameters of enzymatic and non-enzymatic antioxidant defense system and the oxidative modification of lipids. The effect of paracetamol on acute liver injury was also monitored. All analyzed basil extracts showed the presence of the large number of phenolic compounds from the class of phenolic acids and flavonoids. Chlorogenic, p-hydroxybenzoic, caffeic, vanillic, ferulic, rosmarinic, and cinnamic acid, as well as apigenin, quercetin, naringenin and rutin were detected and quantified. The application of basil extracts and the sodium salt of monoketocholic acid, either alone or in combination, and also in pharmaceutical technological formulation of microvesicles has shown a reduction in damage during oxidative stress, then a significant antidiabetic activity, favorable effect on the lipid profile and protective effect on the liver and kidney function. Based on these results it can be concluded that basil extract contains significant amounts of phenolic compounds responsible for antioxidant activity. The aqueous extract of basil, alone or in combination with monoketocholic acid, and in the form of the pharmaceutical technological formulations lowers glucose levels in the blood, accelerates the restoration of &beta; cells, has a favorable effect on the lipid status and does not lead to toxic changes in the liver and kidneys in experimental animals.</p>

<p>Rod Salix pripada porodici Salicaceae i obuhvata oko 450 vrsta &scaron;irom sveta, od kojih u Republici Srbiji raste oko 18. Kora vrbe ispoljava antiinflamatorno, antireumatsko, analgetičko i antipiretičko delovanje sinergističkim efektom njenih glavnih aktivnih supstanci &ndash; glikozida salicina, fenolnih i flavonoidnih jedinjenja. Ciljevi ovog rada bili su ispitivanje uticaja klasične i savremenih metoda ekstrakcije na hemijski sastav i biolo&scaron;ke aktivnosti ekstrakata različitih vrsta vrba, utvrđivanje razlika u sadržaju aktivnih komponenti i biolo&scaron;kom potencijalu između ekstrakata kore i ekstrakata lista iste vrste vrbe, kao i utvrđivanje razlika u sadržaju sekundarnih metabolita i antioksidantnoj aktivnosti kod ekstrakata različitih vrsta vrba. Ispitivanja su uključivala analizu 92 ekstrakta kore i lista osam predstavnika roda Salix, dobijenih različitim metodama (maceracija, ultrazvučna i mikrotalasna ekstrakcija) i pri različitim uslovima ekstrakcije (rastvarač, vreme, veličina čestica). Sadržaj ukupnih fenola i flavonoida, kao i antioksidativna aktivnost određeni su spektrofotometrijski. Hemijska karakterizacija ekstrakata vr&scaron;ena je primenom visokoefikasne tečne hromatografije (HPLC). Metodom in silico molekularnog dokinga ispitan je inhibitorni potencijal sastojaka ekstrakata prema enzimima ciklooksigenaze (COX-1 i COX-2) i acetilholinesterazi (AChE). Savremenim metodama ekstrakcije izolovan je veći broj pojedinačnih komponenata u najvećoj koncentraciji i dobijeni su ekstrakti sa jačim potencijalom neutralizacije hidroksilnih radikala. Klasična metoda maceracije 70% etanolom (v/v) bila je pogodnija za dobijanje ekstrakata sa jačim antioksidativnim potencijalom prema DPPH radikalu kod gotovo svih ispitivanih Salix vrsta. Kod većine Salix vrsta jači antioksidantni potencijal prema DPPH radikalu ispoljili su ekstrakti kore. Kod polovine ispitivanih vrsta ekstrakti lista su inhibisali hidroksilne radikale u većoj meri. Veći sadržaj pojedinačnih jedinjenja u ekstraktima kore ili lista zavisio je od same vrste vrbe. Najjaču antioksidantu aktivnost ispoljila je vrsta S. alba, dok je u pogledu hemijskog sastava najraznovrsnija vrsta bila S. fragilis. In silico analizom metodom molekularnog dokinga utvrđen je jak inhibitorni potencijal flavonoidnih jedinjenja kvercetina, naringenina i epikatehina, kao i hlorogenske kiseline među fenolnim kiselinama prema enzimima COX-1 i COX-2. Epikatehin, salicin i hlorogenska kiselina ispoljili su značajno inhibitorno delovanje na enzim AChE u doking studijama. Dobijeni rezultati ukazuju da pored kore i list vrbe ima veliki potencijal primene kao izvor značajnih fitokomponenata, kao i na mogućnost eksploatacije vrsta vrba koje nisu u komercijalnoj upotrebi kao lekovitih sirovina za izolovanje antioksidanasa i farmakolo&scaron;ki aktivnih supstanci.</p> / <p>The genus Salix belongs to the family Salicaceae and comprises about 450 species worldwide, out of which 18 grow in the Republic of Serbia. Willow bark exhibits anti-inflammatory, anti-rheumatic, analgesic and antipyretic effects through synergistic action of its main ingredients -glycosidesalicin, phenolic and flavonoid compounds. Aims of this paper were to analyse the impact of classical and modern extraction methods on chemical composition and biological activities of extracts of different willow species, determine the differences in the amount of active compounds and biological potential between bark and leaf extracts within the same willow species, as well as between extracts of various species. The analysis included 92 bark and leaf extracts of eight species from the genus Salix, obtained by different extraction methods (maceration, ulrasound and microwave assisted extraction) and conditions (solvent, time, particle size). Total phenolics and flavonoids content, as well as antioxidant activity were determined spectrophotometrically. Chemical characterization was carried out by high performance liquid chromatography (HPLC). Cyclooxygenase (COX-1 and COX-2) and acetylcholine esterase (AChE) inhibitory potential of compounds isolated from the extracts was examined by in silico method of molecular docking. Greater number of individual components in highest concentration, as well as exracts with stronger hydroxyl radical scavenging potential were obtained by modern extraction methods. Classical method of maceration with 70% ethanol (v/v) was more suitable for obtaining extracts with higher DPPH radical scavenging activity in the vast majority of tested species. In half of the analysed species leaf extracts inhibited hydroxyl radicals more than bark. Higher amount of individual compounds in bark or leaf extracts depended on the species itself. S. alba exhibited the strongest antioxidant activity, whereas S. fragilis had the most diverse chemical composition. Strong COX-1 and COX-2 inhibitory potential of flavonoids quercetin, naringenin and epicatechin, as well as chlorogenic acid among phenolic acids was determined by in silico molecular docking analysis. Molecular docking studies also demonstrated the inhibitory activity of epicatechin, salicin and chlorogenic acid toward AChE. The obtained results indicate that not only bark, but willow leaf as well could be used as source of significant phytochemicals and also the possibility of exploitation of willow species that are not commercially used as medicinal raw material for isolation of antioxidants and pharmacologycally active substances.</p>

<p>Ispitani su hemijski sastav i biolo&scaron;ka aktivnost &scaron;est samoniklih vrsta iz tribusa Scandiceae<br />(familija Apiaceae): Anthriscus sylvestris, Anthriscus cerefolium, Chaerophyllum bulbosum,<br />Chaerophyllum hirsutum, Chaerophyllum temulentum i Scandix pecten-veneris. LC-MS-MS<br />analizom ekstrakata identifikovano je vi&scaron;e desetina sekundarnih biomolekula iz klasa flavonoida,<br />fenilpropenskih kiselina, lignana i kumarina. GC-MS analiza pružila je uvid u sastav volatilnih<br />komponenti i njihov hemosistematski značaj. Potvrđeno je da sve ispitivane vrste imaju umereno<br />antioksidantno, antiinflamatorno i antiproliferativno dejstvo.</p> / <p> Chemical composition and biological activity of six wild-growing species from<br /> Scandiceae tribe (Apiaceae family) &ndash; Anthriscus sylvestris, Anthriscus cerefolium, Chaerophyllum<br /> bulbosum, Chaerophyllum hirsutum, Chaerophyllum temulentum and Scandix pecten-veneris &ndash; was<br /> examined. By LC-MS-MS analysis, a large number of secondary biomolecules was identified in<br /> extracts, including flavonoids, phenylpropenic acids, lignans and coumarins. GC-MS analysis<br /> provided insight into volatile components composition and chemosystematic significance. All<br /> investigated species exhibited moderate antioxidant, anti-inflammatory and antiproliferative<br /> activity.</p>

<p>&Scaron;tavelj (Rumex crispus, Polygonaceae) je vi&scaron;egodi&scaron;nja zeljasta biljka, koja predstavlja bogat izvor fenolnih komponenti. Iako se smatra invazivnim korovom, mlado li&scaron;će &scaron;tavelja je jestivo i često se koristi kao salata. Dalje, upotreba plodova &scaron;tavelja opisana je u srpskoj i turskoj narodnoj medicini u lečenju gastrointestinalnih tegoba. Cilj ovog rada bio je procena in vitro i in vivo antioksidantne/prooksidantne i citotoksične aktivnosti, i određivanje eventualnog in vitro antiinflamatornog efekta vodenog ekstrakta ploda Rumex crispus. Ukupan sadržaj flavonoida određen je spektrofotometrijskom metodom. Kvalifikacija i kvantifikacija flavonoida potvrđena je visokoefikasnom tečnom hromatografijom (HPLC). Antioksidantna aktivnost vodenog ekstrakta ploda &scaron;tavelja procenjena je na osnovu in vitro testova: Ferric-reducing antioxidant power (FRAP), sposobnosti ekstrakta da neutrali&scaron;e slobodne radikale NO&bull;, OH&bull; i DPPH&bull; i uticaja na lipidnu peroksidaciju u lipozomima. Citotoksičnost ispitivanog ekstrakta je određena in vitro na tumorskim ćelijskim linijama: humani karcinom cerviksa (HeLa), adenokarcinom (HT-29) i adenokarcinom dojke (MCF7). Takođe, moguća in vivo hepatoprotektivna i antioksidantna svojstva ekstrakta određena su kod oksidativnog stresa izazvanog CCl4 kod eksperimentalnih životinja. Pored toga, proverena je hipoteza u kojoj testiran ekstrakt pokazuje in vivo antiproliferativnu aktivnost kod Ehrlich-ovih (EAC) i Hepatoma AS30D ćelija, merenjem zapremine ascitesa, procenta vijabilnih ćelija i nivoa nekoliko antioksidantnih enzima. Optimizovan in vitro test za određivanje potencijala inhibicije ciklooksigenaze-1 (COX-1) i 12-lipooksigenaze (12-LOX) preduzet je u svrhu procene antiinflamatornog efekta vodenog ekstrakta ploda R. crispus. HPLC analiza otkrila je da je mikvelianin najdominantniji flavonoidni konstituent ekstrakta. Testirani ekstrakt pokazao je potencijalnu antioksidantnu aktivnost rezultujući velikom moći u neutralizaciji slobodnih radikala, i sposobno&scaron;ću da smanji lipidnu peroksidaciju u lipozomima. Rezultati su ukazali na tkivno-selektivnu citotoksičnost ekstrakta ploda R. crispus in vitro. Najizraženija antitumorska aktivnost primećena je prema HeLa i MCF7 ćelijskim linijama. Podaci sugeri&scaron;u da bi se ispitivani ekstrakt mogao smatrati potencijalnim in vivo hepatoprotektivnim i antioksidantnim agensom, sprečavajući oksidativna o&scaron;tećenja jetre. S druge strane, pomenuti ekstrakt može pokazati in vivo prooksidantna svojstva, uzrokujući oksidativni stres u maligno transformisanim EAC i AS30D ćelijama i smanjujući zapreminu ascitesa i udeo vijabilnih ćelija, u poređenju sa kontrolnom grupom. Promene u aktivnosti antioksidantnih enzima su verovatno posledica indukovanog oksidativnog stresa u EAC i AS30D ćelijama, naročito kod pretretiranih životinja. Vodeni ekstrakt ploda &scaron;tavelja pokazao je COX-1, kao i 12-LOX inhibitornu aktivnost, navodeći da bi ispitivani ekstrakt mogao biti antiinflamatorni agens. Vodeni ekstrakt ploda R. crispus ima potencijalnu antioksidantnu, citotoksičnu i antiinflamatornu aktivnost. Ispoljavanje prooksidantnih svojstava predstavlja mogući mehanizam antiproliferativnog efekta ekstrakta.</p> / <p>Curly dock (Rumex crispus, Polygonaceae) is a wild perennial herbaceous plant, which products are described as a rich source of phenolic compounds. Apart from being considered a seriously invasive weed, young leaves of curly dock are edible and often used as salad. Furthermore, the use of its fruits has been described in Serbian and Turkish traditional medicine against stomach complaints. The objectives of this study were to evaluate in vitro and in vivo antioxidant/prooxidant and cytotoxic activities, and to determine an eventual in vitro anti-inflammatory effect of the aqueous extract of Rumex crispus fruits. Total flavonoid content was determined by spectrophotometric method. Qualification and quantification of flavonoids were confirmed using High performance liquid chromatography (HPLC). The aqueous extract of curly dock fruits was evaluated for its antioxidant activity by in vitro assays for Ferric-reducing antioxidant power (FRAP), NO&bull;, OH&bull; and DPPH&bull;-free radical scavenging activities and the influence on lipid peroxidation in liposomes. The cytotoxicity of tested extract was examined in vitro in human cervix carcinoma (HeLa), colon adenocarcinoma (HT-29) and breast adenocarcinoma (MCF7). Also, the potential in vivo hepatoprotective and antioxidant properties of investigated extract were determined on CCl4-induced oxidative stress in experimental animals. Furthermore, the hypothesis that the examined extract might show in vivo antiproliferative activity in Ehrlich carcinoma (EAC) and Hepatoma AS30D cells was tested by measuring volume of ascites, percentage of viable cells and level of several antioxidant enzymes. The optimized in vitro test for determination of cyclooxygenase-1 (COX-1) and 12-lipoxygenase (12-LOX) inhibition potency was undertaken in order to estimate an anti-inflammatory effect of aqueous extract of R. crispus fruits. HPLC analysis revealed miquelianin as the most abundant flavonoid constituent of the extract. The tested extract might have an antioxidant activity resulting in scavenging of free radicals and ability to decrease lipid peroxidation in liposomes. The results could indicate tissue-selective cytotoxicity of R. crispus fruit extract in vitro. The most prominent antitumor activity was observed towards HeLa and MCF7 cell lines. The data suggested that investigated extract may be considered as potential in vivo hepatoprotective and antioxidant agent due to prevention of the liver injuries induced by oxidative damage. On the other hand, mentioned extract could exhibit in vivo prooxidant property, causing the oxidative stress in malignant transformed EAC and AS30D cells and reducing volume of ascites and percentage of viable cells, in comparison with control group. Changes in activities of antioxidant enzymes might be the results of induced oxidative stress in EAC and AS30D cells, especially in the pretreated animals. The aqueous extract of curly dock fruits showed COX-1, as well as 12-LOX inhibitory activity, suggesting that tested extract might be an anti-inflammatory agent. It could be concluded that aqueous fruit extract of R. crispus might have antioxidant, cytotoxic and anti-inflammatory activities. The prooxidant properties of examined extract could be the mechanism of potential antiproliferative effect of extract.</p>

<p>U ovom radu je izvr&scaron;eno kvantitativno&nbsp;određivanje i identifikacija biljnih fenola i&nbsp;<br />flavonoida etanolih ekstrakata ploda per&scaron;una&nbsp;(<em>Petroselini fructus</em>), kore kru&scaron;ine (<em>Frangulae&nbsp;cortex</em>), lista pitome nane (<em>Mentha piperitae&nbsp;folium</em>), ploda kima (<em>Carvi fructus</em>) i lista breze&nbsp;(<em>Betulae folium</em>), kao i komercijalnog preparata Vitalplant^&reg;((<em>Frangulae cortex</em>(35 %),&nbsp; <em>Mentha&nbsp;piperitae folium</em>(20%),&nbsp; <em>Carvi fructus</em>(20 %),&nbsp;<em>Petroselini fructus&nbsp;</em>(25 %)). U nastavku&nbsp;istraživanja ispitivana je antioksidantna aktivnosti etanolih ekstrakata primenom&nbsp;direktnih, ESR &ldquo;spin trap&ldquo; spektroskopskih&nbsp;metoda određivanja &ldquo;skevindžer&ldquo; aktivnosti na&nbsp;superoksid anjon i &nbsp;hidroksil radikale i&nbsp;indirektnih, spektrofotometrijskih testova za&nbsp;određivanje skevindžer&ldquo; aktivnosti na DPPH˙&nbsp;radikale, redoks potencijala i helatacione&nbsp;aktivnosti, kao i antioksidantne aktivnosti u&nbsp;sistemu&nbsp; &beta;-karoten-linolna kiselina. Osim navedenog, deo istraživanja je posvećen&nbsp;ispitivanju termičke stabilnosti ekstrakata&nbsp;navedenih biljaka i komercijalnog preparata&nbsp;Vitalplant^&reg;&nbsp;radi sticanja uvida u mogućnost&nbsp;njihove primene u &nbsp;pekarskim proizvodima. U&nbsp;poslednjoj fazi rada, izvr&scaron;eno je određivanje sposobnosti pulvisa i etanolnog ekstrakta komercijalnog &nbsp;preparata Vitalplant^&reg; da inhibiraju oksidaciju lipida u keksu, primenom spektrofotometrijskih testova &ldquo;skevindžer&ldquo; aktivnosti &nbsp;na DPPH˙ radikale i MDA testa. Ispitane biljne sirovine, kao i komercijalni preparat Vitalplant^&reg;&nbsp;su bogat izvor jedinjenja iz klase biljnih fenola. Sve ispitane biljne droge poseduju antioksidantnu aktivnost, koja se značajno ne menja usled termičkog tretmana. Dodatak biljne me&scaron;avine Vitalplant^&reg;&nbsp;, upravo proporcionalno njenom sadržaju, &nbsp;dovodi do povećanja antioksidantne aktivnosti keksa i smanjenja stepena lipidne peroksidacije.&nbsp;</p> / <p>In this paper, quantitative determination and identification of plant phenolics and flavonoids of ethanolic extracts obtained from parsley fruit (<em>Petroselini fructus</em>), buckthorn bark (<em>Frangulae cortex</em>), mint leaves (<em>Mentha piperitae folium</em>), caraway fruit&nbsp; (<em>Carvi fructus</em>), birch leaves (<em>Betulae folium</em>), as well as from commercial preparation Vitalplant&reg; ((<em>Frangulae cortex</em>(35 %), <em>Mentha piperitae folium</em>&nbsp;(20%), <em>Carvi fructus</em>(20 %), <em>Petroselini fructus&nbsp;</em>(25 %)) was performed. In addition, antioxidant&nbsp; activity of ethanolic extracts was tested, by applying direct, ESR &ldquo;spin trap&ldquo; &nbsp;spectroscopic methods for the determination of scavenging activity on superoxide anion and hydroxyl radicals, and indirect, spectrophotometric methods for the determination of DPPH˙ radical scavenging activity, reducing power, chelating activity and antioxidant activity in&nbsp; &beta;-carotene-linoleic acid model system. Moreover, thermal stability of the ethanolic extracts was tested in order to get insight into possible application of the extracts in bakery products. Finally, the ability of the commercial preparation Vitalplant&reg;, in the form of powder and extract, to inhibit oxidative changes of cookies was tested by applying spectrophotometric DPPH radical scavenging and MDA tests.</p><p>According to obtained results, investigated plant samples, including commercial preparation Vitalplant&reg;, are rich source of plant phenolics. Investigated plant drugs possess antioxidant activity, which is not significantly changed after the thermal treatment. Cookie supplementation with commercial preparation Vitalplant&reg; results in&nbsp; better oxidative stability of lipids and enhanced antioxidant activity of the cookies.</p>

<p>U ovoj doktorskoj disertaciji ispitan je hemijski sastav i biolo&scaron;ke aktivnosti<br />ekstrakata deset&nbsp; samoniklih&nbsp; taksona roda&nbsp; <em>Allium</em>&nbsp; sect.&nbsp;<em> Codonoprasum:&nbsp; A. carinatum</em>&nbsp;subsp. <em>pulchellum,&nbsp; A. carinatum&nbsp;</em> subsp. <em>carinatum,&nbsp; A. fuscum&nbsp;</em> var. <em>gracile,&nbsp; A. fuscum&nbsp;</em> var.&nbsp;<em>fuscum,&nbsp; A. flavum</em> subsp. <em>flavum,&nbsp; A. melanantherum,&nbsp; A. paniculatum&nbsp;</em> subsp.&nbsp;<em> marginatum,&nbsp;A. pallens</em>&nbsp; subsp. <em>tenuiflorum,&nbsp; A. oleraceum&nbsp; </em>i&nbsp;<em> A. rhodopeum</em>, sakupljenih na&nbsp; 27 lokaliteta&nbsp;u Srbiji. Cilj rada bio je da se dobiju podaci o sadržaju biolo&scaron;ki aktivnih jedinjenja u&nbsp;ovim, do sada veoma malo ispitanim vrstama roda&nbsp; <em>Allium</em>, i utvrdi njihova potencijalna&nbsp;lekovita vrednost.</p><p>Analiza hemijskog sastava obuhvatila je: analizu volatilnih komponenti svežih&nbsp;<br />lukovica primenom headspace GC-MS tehnike, kvalitativnu analizu metanolnih &nbsp;<br />ekstrakata primenom tečnohromatografskih metoda (LC-DAD-MS i LC-MS-MS), &nbsp;<br />kvantitativnu analizu odabranih fenolnih jedinjenja LC-MS-MS tehnikom,&nbsp; određivanje&nbsp;sadržaja ukupnih&nbsp; monomernih&nbsp; antocijana&nbsp; i određivanje aktivnosti aliinaze. Ispitivanja&nbsp;biolo&scaron;kih aktivnosti ekstrakata obuhvatila su: određivanje antioksidantne,&nbsp;antiinflamatorne, antimikrobne, antimutagene i genotoksične aktivnosti, kao i ispitivanje&nbsp;uticaja na rast zdravih i tumorskih ćelija i sposobnosti indukcije ćelijske smrti.&nbsp;</p><p>Sumiranjem dobijenih rezultata može se zaključiti da ispitivani predstavnici roda&nbsp;<br /><em>Allium&nbsp;</em> sect.&nbsp; <em>Codonoprasum&nbsp;</em> predstavljaju bogate izvore biolo&scaron;ki aktivnih jedinjenja sa&nbsp;&scaron;irokim spektrom biolo&scaron;kih aktivnosti. Sa hemotaksonomskog aspekta značajno je da se&nbsp;dimetil-disulfid&nbsp; izdvaja kao&nbsp; najdominantnija&nbsp; i često jedina&nbsp; isparljiva komponenta, da&nbsp;ekstrakti većine vrsta&nbsp; sadrže veliku količinu flavonoida (prvenstveno derivata&nbsp;kvercetina), da se vrste&nbsp;<em> A. pallens&nbsp;</em> i&nbsp; <em>A. oleraceum</em>&nbsp; izdvajaju od ostalih po tome &scaron;to ne&nbsp;sadrže rutin&nbsp; a sadrže&nbsp; hiperozid,&nbsp; da je vrsta&nbsp;<em> A. rhodopeum</em>&nbsp; siroma&scaron;na fenolnim&nbsp;jedinjenjima i da su sve vrste, osim vrste&nbsp; <em>A.&nbsp; flavum</em>, bogate antocijanima. Aktivnost&nbsp;aliinaze je visoka u svim ispitivanim vrstama. Većina ispitivanih ekstrakata, izuzev&nbsp;ekstrakata vrsta&nbsp; <em>A. carinatum&nbsp;</em>i <em>A. melanantherum</em>, pokazala je izraženu antioksidantnu&nbsp;aktivnost, dok su ekstrakti vrsta <em>A. flavum</em>, <em>A. rhodopeum</em>, <em>A. oleraceum </em>i <em>A. paniculatum&nbsp;</em>snažni antiinflamatorni agensi. Ekstrakti ispitivanih predstavnika sect.&nbsp;<em> Codonoprasum&nbsp;</em>nisu pokazali antimikrobnu i antimutagenu aktivnost. Takođe, ovi ekstrakti nisu ispoljili&nbsp;genotoksični efekat na ćelije zdravog tkiva (izuzev slabog genotoksičnog efekta ekstrakta&nbsp;nadzemnih delova&nbsp;<em> A. flavum</em>), &scaron;to ukazuje na bezbednost upotrebe vrsta sect.&nbsp;<em>Codonoprasum&nbsp;</em> kao hrane ili u obliku lekova. Ekstrakti celih biljaka&nbsp;<em>A. paniculatum</em> i <em>A.&nbsp;rhodopeum</em>, kao i ekstrakt nadzemnih delova &nbsp;<em>A. melanantherum</em>&nbsp; pokazali su snažnu&nbsp;antiproliferativnu aktivnost sa povoljnim ne-tumor/tumor koeficijentima i indukovali&nbsp;apoptozu u tumorskim ćelijama, iz čega se može zaključiti da imaju visok potencijal&nbsp;primene u antitumorskoj terapiji.&nbsp;</p> / <p>In the present doctoral thesis the chemical composition and biological activities&nbsp; of&nbsp;<br />10 wild growing taxa of genus&nbsp; <em>Allium</em>&nbsp; sect. <em>Codonoprasum</em>&nbsp; (<em>A. carinatum&nbsp;</em> subsp.&nbsp;<br /><em>pulchellum,&nbsp; A. carinatum&nbsp;</em> subsp.&nbsp; <em>carinatum,&nbsp; A. fuscum&nbsp;</em> var.&nbsp; <em>gracile,&nbsp; A. fuscum</em>&nbsp; var.&nbsp;<br /><em>fuscum,&nbsp; A. flavum&nbsp;</em> subsp.&nbsp; <em>flavum,&nbsp; A. melanantherum,&nbsp; A. oleraceum,&nbsp; A. paniculatum</em><br />subsp. <em>marginatum, A. pallens</em> subsp. <em>tenuiflorum</em> and <em>A. rhodopeum</em>) were investigated.&nbsp;The samples were&nbsp; collected from 27 locations in Serbia. The aim of the study was to&nbsp;obtain data on the content of biologically active compounds in extracts of &nbsp;these&nbsp;unexplored species of the genus Allium and to determine their potential medicinal value.</p><p>Phytochemical caracterisation included: headspace GC-MS analysis of fresh bulb&nbsp;<br />volatiles, LC-DAD-MS and LC-MS-MS qualitative analysis of methanol extracts, LC-MS-MS quantitative analysis of 44 selected phenolic compounds in methanol extracts,&nbsp;determination of total monomeric anthocyanins content and alliinase activity. In order to&nbsp;assess the biological potential of methanol extracts, the antioxidant, anti-inflammatory,&nbsp;antimicrobial, antimutagenic, genotoxic and antiproliferative activities of &nbsp;the extracts&nbsp;were studied.</p><p>Summing up all the results obtained, it can be concluded that species of genus&nbsp;<em>Allium&nbsp;</em> sect.&nbsp; <em>Codonoprasum&nbsp;</em> are rich sources of biologically active compounds with a&nbsp;broad spectrum of biological activities. Dimethyl disulfide is the most dominant and&nbsp;often the only volatile component of most species, which is significant from&nbsp;chemotaxonomic point of view. The methanol extracts of investigated species contain&nbsp;high levels of flavonoids (primarily quercetin derivatives). Specificity of&nbsp; <em>A. pallens</em>&nbsp; and&nbsp;<em>A. &nbsp;oleraceum</em>&nbsp; extracts is that they do not contain rutin, but contain hyperoside, while&nbsp;small quantity of phenolic compounds&nbsp; is characteristic for&nbsp; <em>A. rhodopeum</em>&nbsp; extract. All&nbsp;investigated species, except of <em>A. flavum</em>, are rich in anthocyanins. Alliinase activity was&nbsp;high in&nbsp; all examined species. Most of the extracts, except&nbsp; <em>A. carinatum</em>&nbsp; and&nbsp; <em>A.&nbsp;melanantherum</em>&nbsp; extracts, express considerable antioxidant activity, &nbsp;while extracts of&nbsp;<em> A.&nbsp;flavum</em>,&nbsp; <em>A. rhodopeum</em>,&nbsp; <em>A. paniculatum</em>&nbsp; and&nbsp; <em>A. oleraceum&nbsp;</em> are potent anti-inflammatory&nbsp;agents. The investigated&nbsp; <em>Allium</em>&nbsp; extracts did not show antimicrobial and antimutagenic&nbsp;activity. Also, the extracts did not express genotoxic effect on healthy tissue cells&nbsp;(except the weak genotoxic effects of aerial parts extract of&nbsp; <em>A. flavum</em>), indicating that&nbsp;the use&nbsp; of these species as a food or as a drug is safe. Whole plant extracts of&nbsp; <em>A.&nbsp;paniculatum</em>&nbsp; and&nbsp;<em> A. rhodopeum</em>, as well as aerial parts extract of&nbsp;<em> A. melanantherum</em>&nbsp;showed strong antiproliferative activity (with a favorable &nbsp;non-tumor/tumor ratios) and&nbsp;induced apoptosis in tumor cells, suggesting that these plants have a high potential for&nbsp;application in antitumor therapy.</p>