The time- and dose-dependent effects of wortmannin on transepithelial electrical resistance (R(te)) and forskolin-stimulated chloride secretion in T84 monolayer cultures were studied. In both instances, maximal effects developed over 2 h and were stable thereafter. Inhibition of forskolin-stimulated chloride secretion, as measured by the short-circuit current (I(SC)) technique, had an IC50 of 200-500 nM, which is 100-fold higher than for inhibition of phosphatidylinositol 3-kinase (PI3K), but similar to the IC50 for inhibition of myosin light chain kinase (MLCK) and mitogen-activated protein kinases (MAPK). Previous work demonstrated that 500 nM wortmannin did not inhibit the cAMP activation of apical membrane chloride channels. We show here that 500 nM wortmannin has no affect on basolateral Na/K/2Cl-cotransporter activity, but inhibits basolateral membrane Na/K-ATPase activity significantly. The MLCK inhibitors ML-7 and KT5926 were without affect on forskolin-stimulated I(SC). Similarly, the p38- and MEK-specific MAPK inhibitors SB203580 and PD98059 did not reduce forskolin-stimulated I(SC). In contrast, the non-specific MAPK inhibitor apigenin reduced forskolin-stimulated I(SC) and basolateral membrane Na/K-ATPase activity similar to wortmannin. In isolated membranes from T84 cells, wortmannin did not inhibit Na/K-ATPase enzymatic activity directly. We conclude that one or more MAPK may regulate the functional expression of basolateral membrane Na/K-ATPase by controlling the abundance of enzyme molecules in the plasma membrane.

<p>У оквиру ове докторске дисертације изведено је<br />испитивање различитих екстракционих поступака за<br />изоловање апигенина из цвета камилице, као и евалуација<br />биолошке активности добијених екстраката. Полазни<br />биљни материјал сачињавале су две групе латица<br />камилице: ферментисане и неферментисане (нативне).<br />Екстракција ферментисаних цветова је извођена применом<br />ултразвучне екстракције користећи етанол као екстрагенс,<br />а добијени екстракти су се одликовали изузетно високим<br />садржајем апигенина. Оптимизација екстракције је била<br />изведена применом методе одзивне површине. Применом<br />електрон-спин резонанце испитана је антирадикалска<br />активност екстраката. Додатно, фармаколошка вредност<br />добијених екстраката је потврђена и одређивањем њиховог<br />антимикробног и антипролиферативног потенцијала.<br />Нативни цветови камилице су екстраховани применом<br />различитих екстаркционих техника: микроталасне,<br />ултразвучне, Soxhlet екстракције као и екстракције<br />субкритичном водом. Eкстрaкција водом у субкритичном<br />стању се показала супериорнијом у односу на све остале<br />технике у погледу садржаја укупних фенола и флавоноида.<br />У циљу добијања екстраката са максималним садржајем<br />апигенина изведена је оптимизација овог екстракционог<br />процеса. Изоловање чистог апигенина је изведено из<br />екстракта добијеног под оптималним екстракцијом<br />условима (однос дрога:растварач 1:30, брзина мешања 3 Hz,<br />притисак 45 bar, температура 115&ordm;C, време 30 мин,<br />концентрација модификатора 0,001 М) применом поступка<br />колонске хроматографије на стубу полиамида. Хемијски<br />профил као и садржај појединачних полифенолних<br />компонената у екстрактима добијеним на различитим<br />притисцима, температурама и уз присуство модификатора<br />различитих концентрација одређен је применом UHPLCDAD-<br />HESI-MS/MS. У свим анализираним екстрактима<br />детектован је велики број полифенолних компонената, док<br />је апигенин у свима био доминантно једињење. Садржај<br />апигенина у екстракту добијеном под оптималним<br />екстракционим условима је износио 1.700,34 mg/kg.<br />Применом седам различитих тестова извршена је<br />евалуација антиоксидативног и антирадикалског<br />потенцијала екстраката. Антимикробни потенцијал<br />екстраката је одређен за осам различитих микробних<br />линија. in vitro тестовима испитана је способност<br />инхибиције &alpha;-амилазе, &alpha;-глукозидазе и тирозиназе.<br />Деловањем на раст три хистолошки различите ћелијске<br />линије, испитана је антипролиферативна активност<br />екстраката добијених субкритичном водом.<br />Антимотилитетна активност обе групе екстраката<br />(ферментисаних и неферментисаних цветова) одређена је у<br />in vitro условима.</p> / <p>U okviru ove doktorske disertacije izvedeno je<br />ispitivanje različitih ekstrakcionih postupaka za<br />izolovanje apigenina iz cveta kamilice, kao i evaluacija<br />biološke aktivnosti dobijenih ekstrakata. Polazni<br />biljni materijal sačinjavale su dve grupe latica<br />kamilice: fermentisane i nefermentisane (nativne).<br />Ekstrakcija fermentisanih cvetova je izvođena primenom<br />ultrazvučne ekstrakcije koristeći etanol kao ekstragens,<br />a dobijeni ekstrakti su se odlikovali izuzetno visokim<br />sadržajem apigenina. Optimizacija ekstrakcije je bila<br />izvedena primenom metode odzivne površine. Primenom<br />elektron-spin rezonance ispitana je antiradikalska<br />aktivnost ekstrakata. Dodatno, farmakološka vrednost<br />dobijenih ekstrakata je potvrđena i određivanjem njihovog<br />antimikrobnog i antiproliferativnog potencijala.<br />Nativni cvetovi kamilice su ekstrahovani primenom<br />različitih ekstarkcionih tehnika: mikrotalasne,<br />ultrazvučne, Soxhlet ekstrakcije kao i ekstrakcije<br />subkritičnom vodom. Ekstrakcija vodom u subkritičnom<br />stanju se pokazala superiornijom u odnosu na sve ostale<br />tehnike u pogledu sadržaja ukupnih fenola i flavonoida.<br />U cilju dobijanja ekstrakata sa maksimalnim sadržajem<br />apigenina izvedena je optimizacija ovog ekstrakcionog<br />procesa. Izolovanje čistog apigenina je izvedeno iz<br />ekstrakta dobijenog pod optimalnim ekstrakcijom<br />uslovima (odnos droga:rastvarač 1:30, brzina mešanja 3 Hz,<br />pritisak 45 bar, temperatura 115&ordm;C, vreme 30 min,<br />koncentracija modifikatora 0,001 M) primenom postupka<br />kolonske hromatografije na stubu poliamida. Hemijski<br />profil kao i sadržaj pojedinačnih polifenolnih<br />komponenata u ekstraktima dobijenim na različitim<br />pritiscima, temperaturama i uz prisustvo modifikatora<br />različitih koncentracija određen je primenom UHPLCDAD-<br />HESI-MS/MS. U svim analiziranim ekstraktima<br />detektovan je veliki broj polifenolnih komponenata, dok<br />je apigenin u svima bio dominantno jedinjenje. Sadržaj<br />apigenina u ekstraktu dobijenom pod optimalnim<br />ekstrakcionim uslovima je iznosio 1.700,34 mg/kg.<br />Primenom sedam različitih testova izvršena je<br />evaluacija antioksidativnog i antiradikalskog<br />potencijala ekstrakata. Antimikrobni potencijal<br />ekstrakata je određen za osam različitih mikrobnih<br />linija. in vitro testovima ispitana je sposobnost<br />inhibicije &alpha;-amilaze, &alpha;-glukozidaze i tirozinaze.<br />Delovanjem na rast tri histološki različite ćelijske<br />linije, ispitana je antiproliferativna aktivnost<br />ekstrakata dobijenih subkritičnom vodom.<br />Antimotilitetna aktivnost obe grupe ekstrakata<br />(fermentisanih i nefermentisanih cvetova) određena je u<br />in vitro uslovima.</p> / <p>In the frame of this thesis different extraction approaches for<br />apigenin isolation from chamomile ligulate flowers were<br />examined and biological activity of obtained extracts was<br />evaluated. Starting plant samples included fermented and<br />nonfermented (native) flowers.<br />Extraction of fermented flowers was performed by using<br />ultrasound-assisted extraction with ethanol. The concentration<br />of apigenin was high in obtained extracts. Optimization of the<br />extraction procedures was performed by response surface<br />methodology. Antiradical activity of observed extracts was<br />examined by electron-spin resonance spectroscopy.<br />Furthermore, pharmacological potential of obtained extracts<br />was confirmed by determining their antimicrobial and<br />antiproliferative activity.<br />Native chamomile flowers were extracted by different<br />extraction techniques: microwave, ultrasound, Soxhlet and<br />subcritical water extraction. Subcritical water extraction<br />showed to be superior in comparison to other applied techniques<br />in respect to total phenols and flavonoids content. Optimization<br />of the subcritical water extraction was directed to maximization<br />of apigenin content. Isolation of pure apigenin from extracts<br />obtained under optimal extraction conditions (sample-tosolvent<br />ratio 1:30, agitation rate 3 Hz, temperature 115&ordm;C,<br />pressure 45 bar, extraction time 30 min) was performed by<br />preparative chromatography. Chemical profiles and content of<br />individual polyphenolic components in extracts obtained at<br />different pressures, temperatures, and with different<br />concentrations of a modifier was determined by UHPLC-DADHESI-<br />MS/MS. In all analyzed extracts the great number of<br />polyphenolic components was detected while apigenin was the<br />dominant compound in all extracts. Content of apigenin in the<br />extract obtained under optimal extraction condition was<br />1,700.34 mg/kg. Antioxidant and antiradical potential of<br />extracts was evaluated according to different mechanisms.<br />Antimicrobial potential of extracts was determined against eight<br />different microbial strains. Ability of extracts to inhibit &alpha;-<br />amylase, &alpha;-glucosidase and tyrosinase was determined by in<br />vitro assays. Antiproliferative activity of subcritical water<br />extracts was defined by testing their influence on the growth of<br />three histologically different cell lines.<br />Anti-intestinal motility activity of both group of extracts (native<br />and fermented) was determined by in vivo experiments.</p>

<p>Raloksifen je predstavnik selektivnih modulatora estrogenih receptora koji se koristi u terapiji osteoporoze i invazivnog oblika raka dojke u postmenopauzi. Raloksifen se relativno dobro resorbuje iz gastrointestinalnog trakta, ali pri prvom prolasku kroz jetru podleže biotransformaciji u značajnom procentu, &scaron;to je uzrok njegove niske biolo&scaron;ke raspoloživosti. Bioraspoloživost kod ljudi iznosi 2%, a kod Wistar pacova 39%. Različite supstance se koriste da bi se pobolj&scaron;ala bioraspoloživost lekova. Žučne kiseline, kao &scaron;to je deoksiholna kiselina, omogućavaju bolji prolazak kroz biolo&scaron;ke membrane drugim supstancama, te mogu povećati bioraspoloživost lekova. Apigenin je &scaron;iroko rasprostranjeni flavonoid koji inhibi&scaron;e različite metaboličke puteve i na taj način može usporiti metabolizam i eliminaciju i povećati koncentraciju lekova u krvi. Ciljevi ovog istraživanja su bili da se ispita da li apigenin i natrijum-deoksiholat mogu povećati bioraspoloživost raloksifena, njihov uticaj na biohemijske parametre i parametre hemostaze, kao i da se ispita antioksidativni potencijal apigenina. Ispitan je i uticaj apigenina na akutno o&scaron;tećenje jetre usled primene toksične doze paracetamola. U istraživanju su kori&scaron;ćeni zdravi, beli pacovi mu&scaron;kog roda, soja Wistar. U ogledu su ukupno kori&scaron;ćene 84 eksperimentalne životinje. Sva ispitivanja na životinjama je odobrila Etička komisija Univerziteta u Novom Sadu. Raloksifen je primenjen intravenski i per os, dok su natrijum-deoksiholat i apigenin aplikovani peroralno. Uzorci krvi, urina i fecesa su kori&scaron;ćeni za određivanje farmakokinetskih parametara, dok su za određivanje biohemijskih, hemostatskih i parametara oksidativnog stresa kori&scaron;ćeni serum i uzorci jetre laboratorijskih životinja. Pretretman natrijum-deoksiholatom je doveo do smanjenja koncentracije raloksifena u krvi zbog olak&scaron;anog i brzog prodora raloksifena u periferne kompartmane. Time je značajno produženo poluvreme eliminacije i srednje vreme zadržavanja raloksifena i značajno je povećan volumen distribucije raloksifena. Apigenin je doveo do manjeg pada koncentracije raloksifena u prvim satima nakon intravenske primene raloksifena, dok su koncentracije raloksifena bile značajno vi&scaron;e nakon osmog časa od primene leka. Uticaj raloksifena na biohemijske parametre je bio značajno veći nakon intravenske nego nakon peroralne primene. Nakon intravenske primene raloksifena je značajno povećana aktivnost enzima jetre, ALP, ALT, AST i GGT, dok su pokazatelji funkcije bubrega, urea, mokraćna kiselina i kreatinin bili sniženi. U grupama koje su pretretirane natrijum-deoksiholatom i apigeninom vrednosti ovih parametara bile su niže u odnosu na grupu tretiranu samo raloksifenom. Statistički najznačajniji uticaj je imala primena trojne kombinacije, raloksifena, natrijum-deosiholata i apigenina, koja je dovela do značajnog pada aktivnosti enzima jetre, i u odnosu na grupu tretiranu raloksifenom i u odnosu na kontrolnu grupu. Kod životinja tretiranih kombinacijom apigenina i paracetamola pokazatelji toksičnosti su bili značajno niži, naročito vrednosti ALT i ALP, u odnosu na grupu koja je dobijala samo paracetamol. Hepatotoksičnost izazvana toksičnom dozom paracetamola je potvrđena i histopatolo&scaron;kim promenama na jetri, koje nisu primećene u grupi životinja tretiranih kombinacijom apigenina i paracetamola. Ispitivanjem je utvrđeno da apigenin može da spreči paracetamolom indukovano povećanje nivoa MDA, &scaron;to ukazuje da apigenin pozitivno utiče na očuvanje integriteta ćelije. Aktivnost enzima CAT i GR u homogenatima jetre je bila značajno povećana nakon primene toksične doze paracetamola u odnosu na kontrolnu grupu. Aktivnost enzima CAT i GR u grupi tretiranoj kombinacijom apigenina i paracetamola je bila približna vrednostima u kontrolnoj grupi. Na osnovu rezultata istraživanja može se zaključiti da natrijum-deoksiholat i apigenin značajno utiču na farmakokinetiku raloksifena. Primena natrijum-deoksiholata dovela je do pada koncentracije raloksifena u krvi, značajnog prelaska raloksifena iz krvi u periferne kompartmane i povećanja njegovog volumena distribucije, dok je apigenin značajno usporio metabolizam i eliminaciju raloksifena i doveo do njegovog produženog zadržavanja u krvi. Natrijum-deoksiholat i apigenin su pokazali pozitivan uticaj na biohemijske parametre, parametre hemostaze i smanjenje nivoa oksidativnog stresa. Kombinacija natrijum-deoksiholata i apigenina je pokazala sinergistički uticaj na navedene parametre, odnosno dovela je do značajnih promena u odnosu na pojedinačnu primenu ovih supstanci. Rezultati ispitivanja ukazuju na to da apigenin smanjuje stepen lipidne peroksidacije i da dovodi do značajnog povećanja enzimskih antioksidantnih mehanizama odbrane kod pacova kod kojih je hepatotoksičnost indukovana paracetamolom.</p> / <p>Raloxifene is selective estrogen receptor modulator used in treatment of osteoporosis and invasive breast cancer in postmenopausal women. Raloxifene is well absorbed from the gastrointestinal tract, but undergoes extensive first-pass metabolism, which results in very low bioavailability of raloxifene, 2% in humans, and 39% in Wistar rats. Various supstances are used for increasing bioavailability of other drugs. Bile acids, such as deoxycholic acid, promote transport of other supstances through biological membranes, and consequently, may increase their bioavailability. Apigenin is a widespread flavonoid, which inhibits different metabolic pathways. Thus, apigenin can slow down metabolism and elimination of drugs, and raise drug concentration in blood. Aims of this study were to investigate if apigenin and sodium deoxycholate could increase bioavailability of raloxifene, their influence on biochemical and hemostasis parameters, and to investigate antioxidative potential of apigenin. Furthermore, influence of apigenin on acute liver damage after toxic dose of paracetamol was examined. In vivo experiments were performed on 84 laboratory healthy male Wistar rats. All experiments were approved by Ethics Committee of University of Novi Sad. Raloxifene was applied intravenously and per os, while sodium deoxycholate and apigenin were given perorally. Blood, urine and feces samples were used for pharmacokinetic parameters measurement, whereas serum and liver samples were used for evaluation of biochemical, hemostasis and oxidative stress parameters. Pretreatment of sodium deoxycholate led to raloxifene blood concentration decrease due to easier penetration of raloxifene in peripher compartments. As a result, raloxifene half-life and mean residence time were significantly longer and volume of distribution was increased. Apigenin caused lower decrease in raloxifene concentration in first few hours after raloxifene intravenous application, while raloxifene concentrations after apigenin pretreatment were significantlny higher 8 hours after raloxifene application. Influence of raloxifene on biochemical parameters was more significant after intravenous than after per os application. Intravenous application of raloxifene led to increased activity of liver enzymes, ALP, ALT, AST and GGT, while parameters of kidney function, urea, uric acid and creatinine were decreased in comparison to the control group. In experimental groups pretreated with sodium deoxycholate and apigenin these parameters were lower than in the group treated only with raloxifene. Statistically the most significant effects were in the group treated with combination of raloxifene, sodium deoxycholate and apigenin, which caused significant decrease in activity of liver enzymes compared both with raloxifene and control group of animals. In experimental animals treated with combination of apigenin and paracetamol bioindicators of paracetamol toxicity were significantly lower, especially activity of ALT and ALP, in comparison to the group treated only with paracetamol. Hepatotoxicity induced by toxic dose of paracetamol was also confirmed by histopathological alterations in liver, which were not observed in the experimental group treated with combination of apigenin and paracetamol. In this study it was confirmed that apigenin could prevent paracetamol-induced MDA level increase, which suggests that apigenin have positive effects on cell integrity. Activity of CAT and GR in liver homogenates was significantly increased after toxic dose of paracetamol in comparison to the control group, while activity of these enzymes in the group treated with apigenin and paracetamol was similar to values in the control group. Results of this study showed that sodium deoxycholate and apigenin can significantly change pharmacokinetic parameters of raloxifene. Sodium deoxycholate caused signicant decrease in raloxifene blood concentration, extensive distribution from blood to peripheral compartments and increase of raloxifene volume of distribution. Apigenin inhibited metabolism and elimination of raloxifene and thus prolonged half-life and mean residence time of raloxifene. Sodium deoxycholate and apigenin showed positive effects on biochemical and hemostasis parameters and decreased the level oxidative stress. Combination of sodium deoxycholate and apigenin showed synergistic effects on these parameters in comparison to effects of separate application of sodium deoxycholate and apigenin. The result of our study indicates that apigenin inhibits the level of lipid peroxidation and significantly increase the enzyme antioxidant defence mehanisms in paracetamol induced hepatotoxicity in rats.</p>

Le but de cette thèse de doctorat fut de développer des nanoparticules pour la délivrance de deux molécules hydrophobes de faible poids moléculaire, l’apigénine (AG) et un ferrocifène (FcTriOH), comme stratégie innovante pour le traitement du glioblastome(GBM). Dans un premier temps, différents types de nanoparticules, liposomes, nanocapsules lipidiques (LNC), et nanocapsules à base de polymères, furent formulés et comparés en termes de caractéristiques physico-chimiques, de libération en drogue ou encore de toxicité. Les LNCs furent ainsi sélectionnées. Dans un deuxième temps, les LNCs furent fonctionnalisées en surface par un peptide pénétrant (CPP). La concentration de peptide fut augmenté afin d’améliorer significativement l’internalisation des LNCsdans des cellules humaines de GBM. Les mécanismes de macropinocytose et d’endocytose dépendant de la clathrine et de la cavéoline furent observés. De plus, il fut montré que l’internalisation de ces LNCs fonctionnalisées était réduite dans les cellules saines humaines d’astrocyte. L’efficacité biologique des LNCs chargées en AG et chargées en FcTriOH fut évaluée et comparée : le résultat le plus prometteur fut obtenu avec les LNCs chargées en FcTriOH. Une administration intracérébrale des LNCs sur un modèle tumoral murin orthotopique montra une potentielle toxicité et un besoin d’optimiser la dose administrée. Pour finir, les études menées sur un modèle tumoral ectopique murin montrèrent des résultats prometteurs, après une administration parentérale des LNCs chargées en FcTriOH. Ainsi, cette dernière formulation pourrait ouvrir la voie au développement d’une stratégie thérapeutique alternative pour le traitement du GBM. / The aim of this thesis was to develop nanocarriers for efficient delivery of two low molecular weight hydrophobic drugs, apigenin (AG) and a ferrocifen-derivative(FcTriOH) to glioblastoma (GBM) as potential therapeutic strategies. Firstly, two liposomes, a lipid nanocapsule (LNC), and a polymer-based nanocapsule were develope dand compared by their physicochemical characteristics, drug loading capacity, storage stability, stability in biological serum, drug release profiles, complement consumption and toxicity. Due to various advantageous characteristics, the LNCs were selected for further optimization. Secondly, the LNCs were surface functionalized by adsorbing a GBM-targeting cellpenetratingpeptide (CPP). The CPP concentration increased to significantly enhance LNCinternalization in human GBM cells. The uptake mechanisms observed in U87MG cellswere : micropinocytosis, clathrin-dependent and caveolin-dependent endocytosis. Moreover, the optimized CPP-functionalized LNCs were internalized preferentially in theGBM cells compared to normal human astrocytes. Additionally, the in vitro efficacy of the AG-loaded and FcTriOH-loaded LNCs was evaluated. The FcTriOH-loaded LNC-CPP showed the most promising activity with a low IC50 of 0.5 μM against U87MG cells. Intracerebral administration of the LNCs in a murine orthotopic U87MG tumor modelshowed possible toxic effects and the need for dose optimization. Finally, studies inmurine ectopic U87MG tumor model showed promising activity after parenteral administration of the FcTriOH-loaded LNCs. Overall, these results exhibit the promising activity of FcTriOH-loaded LNCs as potential alternative GBM therapy strategy.

Made available in DSpace on 2017-07-10T19:24:23Z (GMT). No. of bitstreams: 1 Tese Francielly_ Torres dos Santos.pdf: 2692597 bytes, checksum: db1b8713e9d8a55de139153f5a86a29e (MD5) Previous issue date: 2016-02-16 / Parsley has been widely traded as a flavoring in Brazil and worldwide. The quality to produce salsa depends on its cropping management. This, this study aimed to evaluate the effect of alternative organic substrates in productivity and nutritional quality of Petroselium crispum. The treatments consisted of five organic compounds obtained by agro-industrial waste composting process of broiler production chain in which they varied the main source of carbon, i.e, waste cotton carding, pruning of ground urban trees, sawdust, bagasse of milled cane and ground napier grass. In order to obtain the organic substrates, it was added 0, 15, 30, 45 and 60% biochar to each of the five organic compounds, a charcoal obtained from burning wood in boilers. The experimental design was completely randomized in a 5x5 factorial design, with four replications. The nutritional quality of salsa was determined by N, P, K evaluation, and antioxidant activity and productivity. Concerning substrates, functional groups and determination of humification degree were evaluated, correlated with parsley dry weight. It was concluded that the use of substrate from Napier grass ground does not favor Petroselium crispum production. Adding biochar associated with organic compost from urban tree pruning whose main source is carbon favors flavonoid content of Diosmetin-apiosilglucoside-isomer and apigenin-malonyl-glucoside in Petroselium crispum. The electrical conductivity of the organic substrate is the main factor that contains Petroselium crispum production while addition of biochar can minimize this effect / A salsa é amplamente comercializada como especiaria no Brasil e no mundo. A produção de salsa com qualidade depende da forma de cultivo. Assim, neste trabalho, objetivou-se avaliar o efeito de substratos orgânicos alternativos na produtividade e qualidade nutricional de salsa graúda portuguesa. Os tratamentos consistiram de cinco compostos orgânicos, obtidos pelo processo de compostagem de resíduos agroindustriais da cadeia produtiva do frango de corte em que se variou a principal fonte de carbono, a saber: resíduos de desfibrilação de algodão, resíduos de poda de árvores urbanas trituradas, serragem, bagaço de cana moído e capim-napier triturado. Para obtenção dos substratos orgânicos, a cada um dos cinco compostos orgânicos, acrescentaram-se 0, 15, 30, 45 e 60% de biochar, um carvão obtido da queima da madeira em caldeiras. O delineamento experimental adotado foi o inteiramente casualizado, em esquema fatorial 5x5, com quatro repetições. A qualidade nutricional da salsa foi determinada pela avaliação de N, P, K, atividade antioxidante e produtividade. Nos substratos, foram avaliados os grupos funcionais e a determinação do grau de humificação, correlacionados com a matéria seca da salsa. Concluiu-se que o uso de substrato originado de capim-napier triturado não favorece à produção de salsa graúda portuguesa. A adição de biochar associado ao composto orgânico, proveniente de poda de árvores urbanas como principal fonte de carbono, favorece o teor de flavonoides de diosmetin-apiosilglucosídeo isômero e apigenina-malonil-glucosídeo na salsa graúda portuguesa. A condutividade elétrica do substrato orgânico é o fator que mais limita a produção de matéria de salsa graúda portuguesa e a adição de biochar pode minimizar tal efeito.

Made available in DSpace on 2017-05-12T14:47:40Z (GMT). No. of bitstreams: 1 Tese Francielly_ Torres dos Santos.pdf: 2692597 bytes, checksum: db1b8713e9d8a55de139153f5a86a29e (MD5) Previous issue date: 2016-02-16 / Parsley has been widely traded as a flavoring in Brazil and worldwide. The quality to produce salsa depends on its cropping management. This, this study aimed to evaluate the effect of alternative organic substrates in productivity and nutritional quality of Petroselium crispum. The treatments consisted of five organic compounds obtained by agro-industrial waste composting process of broiler production chain in which they varied the main source of carbon, i.e, waste cotton carding, pruning of ground urban trees, sawdust, bagasse of milled cane and ground napier grass. In order to obtain the organic substrates, it was added 0, 15, 30, 45 and 60% biochar to each of the five organic compounds, a charcoal obtained from burning wood in boilers. The experimental design was completely randomized in a 5x5 factorial design, with four replications. The nutritional quality of salsa was determined by N, P, K evaluation, and antioxidant activity and productivity. Concerning substrates, functional groups and determination of humification degree were evaluated, correlated with parsley dry weight. It was concluded that the use of substrate from Napier grass ground does not favor Petroselium crispum production. Adding biochar associated with organic compost from urban tree pruning whose main source is carbon favors flavonoid content of Diosmetin-apiosilglucoside-isomer and apigenin-malonyl-glucoside in Petroselium crispum. The electrical conductivity of the organic substrate is the main factor that contains Petroselium crispum production while addition of biochar can minimize this effect / A salsa é amplamente comercializada como especiaria no Brasil e no mundo. A produção de salsa com qualidade depende da forma de cultivo. Assim, neste trabalho, objetivou-se avaliar o efeito de substratos orgânicos alternativos na produtividade e qualidade nutricional de salsa graúda portuguesa. Os tratamentos consistiram de cinco compostos orgânicos, obtidos pelo processo de compostagem de resíduos agroindustriais da cadeia produtiva do frango de corte em que se variou a principal fonte de carbono, a saber: resíduos de desfibrilação de algodão, resíduos de poda de árvores urbanas trituradas, serragem, bagaço de cana moído e capim-napier triturado. Para obtenção dos substratos orgânicos, a cada um dos cinco compostos orgânicos, acrescentaram-se 0, 15, 30, 45 e 60% de biochar, um carvão obtido da queima da madeira em caldeiras. O delineamento experimental adotado foi o inteiramente casualizado, em esquema fatorial 5x5, com quatro repetições. A qualidade nutricional da salsa foi determinada pela avaliação de N, P, K, atividade antioxidante e produtividade. Nos substratos, foram avaliados os grupos funcionais e a determinação do grau de humificação, correlacionados com a matéria seca da salsa. Concluiu-se que o uso de substrato originado de capim-napier triturado não favorece à produção de salsa graúda portuguesa. A adição de biochar associado ao composto orgânico, proveniente de poda de árvores urbanas como principal fonte de carbono, favorece o teor de flavonoides de diosmetin-apiosilglucosídeo isômero e apigenina-malonil-glucosídeo na salsa graúda portuguesa. A condutividade elétrica do substrato orgânico é o fator que mais limita a produção de matéria de salsa graúda portuguesa e a adição de biochar pode minimizar tal efeito.

This work concerns the amount of certain phenols in some of the representatives of families Alliaceae, Lamiaceae and Apiaceae. Phenols in plants are widely represented and highly concentrated. Flavonoids are one of the smallest, yet quite significant classes of phenols. Ingestion of food containing flavonoids can prevent certain diseases such as Arteriosclerosis. cardiovascular and tumor diseases. This work focuses, for their special biological effects, on five flavonoids: Kaempferol, Quercetin, Myricetin, Apigenin and Luteolin. For determining the content of phenols a method of High-performance liquid chromatography (HPLC) was used. Three representative of the family Alliaceae, two representatives of the family Lamiaceae and three representatives of the family Apiaceae were analysed. Only edible parts of the plants were used for the analysis and the amount of phenols was compared in certain plants planted in beddings to the plants planted in greenhouses. At first, the qualitative representantion of phenols was ascertained by the HPLC method. The result is chromatographic profiles, which were then used in calculating the amount of particular phenols. Then the total amounts of kaempferol, quercetin, myricetin, apigenin and luteolin were found out thanks to HPLC method. The biggest amount of total kaempferol was determined in petroselium hortense planted in beddings (588 mg/kg of fresh sample) and in a greenhouse (340 mg/kg of fresh sample). The biggest amount of quercetin was determined in red onion planted in a greenhouse (773 mg/kg of fresh sample) and in ocimum basilicum planted in beddings (535 mg/kg of fresh sample). The biggest amount of apigenin was determined in petroselium hortense planted in a greenhouse (1790 mg/kg of fresh sample) and in petroselium chrispum planted in beddings (3690 mg/kg of fresh sample).

La apigenina (4', 5, 7 trihidroxiflavona), flavonoide presente en distintos vegetales, tiene numerosas características saludables por las cuales elegimos un derivado hidrosoluble, la apigenina potásica, para el presente estudio.Hemos estudiado su actividad frente a la inflamación aguda, al cáncer de próstata y a los daños causados por las radiaciones, tanto ionizantes como no ionizantes (radiación UV), utilizando diversas técnicas, tanto in vitro como en modelos in vivo. Este flavonoide demostró actividad antiinflamatoria, reduciendo la inflamación hasta un 78%. Asimismo, la apigenina potásica mostró actividad quimiopreventiva frente al cáncer de próstata, al reducir la viabilidad y la migración e inducir apoptosis in vitro, y aumentar la supervivencia de animales con tumores. Además, la apigenina potásica presentó efecto geno- y citoprotector frente a la radiación ionizante (radiación &#947; y X), con un factor de protección de un 27-35 % en linfocitos humanos, y de un 50-86 % en dos líneas celulares de próstata. Por último, el flavonoide ha demostrado proteger frente al fotoenvejecimiento causado por la radiación UV, reduciendo la displasia epitelial y la elastosis dérmica, dos marcadores de cáncer cutáneo; además, ha sido posible detectar la apigenina en diversos tejidos, entre ellos el cerebro. / Apigenin (4', 5, 7 trihidroxyflavone), a flavonoid present in different plants, shows some healthy characteristics for which a water soluble derivated, potassium apigenin, was chosen for this study.It has been studied the activity of potassium apigenin against acute inflammation, prostate cancer and the effect of ionising and non-ionising (UV) radiations, using different techniques, both in vitro and in vivo models.This flavonoid showed anti-inflammatory effects, inhibiting the inflammation by up to 78%. It also demonstrated chemopreventive activity against prostate cancer, by reducing the cell viability and migration, inducing apoptosis and increasing the animal survival. Moreover, potassium apigenin showed genoprotector and citoprotector effects against ionising radiation (radiation &#947; and X), with a protection factor of 27-35% in human lymphocytes and of 50-86% in two prostate cell lines. Finally, the flavonoid protected from the photoaging induced by UV radiation, diminishing epithelial dysplasia and dermal elastosis, two markers of skin cancer; furthermore, potassium apigenin was detected in some tissues, brain among them.

[ES] La actual problemática medioambiental está demandando sistemas de producción más sostenibles, como los de la agricultura ecológica. Una buena alternativa para estos sistemas son las variedades tradicionales de pimiento, con rasgos de calidad característicos y de amplia variabilidad genética, fundamental en los programas de mejora, concretamente para la adaptación a la producción ecológica. En la presente tesis se ha estudiado la calidad nutricional y organoléptica del cultivo del pimiento (Capsicum sp.) en los dos estados de madurez comercial y bajo diferentes prácticas culturales (sistemas ecológico y convencional). Se han analizado los principales compuestos nutricionales de los frutos de pimiento: el ácido ascórbico, los compuestos fenólicos y los carotenoides; así como la fracción volátil, y se ha estudiado su variación en una amplia colección varietal. Adicionalmente, se ha estudiado la fertilidad biológica del suelo a través de las actividades enzimáticas de la fosfatasa alcalina y la catalasa en interacción con el cultivo del pimiento bajo los sistemas de cultivo convencional y ecológico. El estudio de la calidad nutricional mostró amplios rangos de variación según el factor genotipo, aunque el estado de maduración y el sistema de cultivo también contribuyeron a las diferencias observadas. En general, los frutos maduros presentaron niveles superiores de compuestos bioactivos, especialmente para el ácido ascórbico. Asimismo, el cultivo ecológico proporcionó, en general, niveles más altos de ácido ascórbico y compuestos fenólicos en estado maduro. La notable interacción genotipo × sistema de cultivo para el contenido en ácido ascórbico y fenoles totales, y en menor intensidad para el contenido en flavonoides individuales, permitió la selección de varias accesiones con alto rendimiento en estos compuestos, bajo condiciones de cultivo ecológico en ambos estados de maduración. Por el contrario, los efectos del sistema de cultivo y la interacción genotipo × sistema de cultivo no resultaron significativos en los niveles de carotenoides, cuya variación se debió principalmente al factor genotipo. El estudio de la composición volátil permitió encontrar amplias diferencias cuantitativas y cualitativas para este factor de calidad organoléptica en los frutos de pimiento, dependiendo principalmente del factor genético y del estado de maduración, mientras que el sistema de cultivo afectó en menor medida y significativamente a algunos compuestos volátiles individuales. Se encontraron un total de 124 y 122 volátiles en estado inmaduro y maduro, respectivamente. Los cambios en el perfil volátil con la maduración fueron evidentes y muy diversos según el genotipo. En general, el sistema de cultivo ecológico favoreció la acumulación de compuestos volátiles en ambos estados de maduración y durante el proceso de maduración. Las actividades enzimáticas fosfatasa alcalina y catalasa aumentaron durante la fase de cultivo, con actividades máximas generales en la fase intermedia, especialmente para la fosfatasa alcalina. El sistema de cultivo tuvo una mayor contribución a la variación, resultado significativo y superior para el sistema convencional en fase intermedia para ambas actividades y estando generalmente seguido del factor genotipo, significativo para la fosfatasa alcalina en fase intermedia. La interacción genotipo × sistema de cultivo permitiría seleccionar variedades, con capacidad para que los suelos acumularan mayores valores de las actividades enzimáticas fosfatasa alcalina y catalasa, en el sistema de cultivo ecológico. En definitiva, los resultados de este trabajo proporcionarán información de especial utilidad para programas de investigación en hortalizas de alto valor añadido y programas de mejora de la calidad en agricultura ecológica, así como para el control de autenticidad de variedades tradicionales. / [CA] L'actual problemàtica mediambiental està demandant sistemes de producció més sostenibles, com els de l'agricultura ecològica. Una bona alternativa per a aquests sistemes són les varietats tradicionals de pebrot, amb trets de qualitat característics i d'àmplia variabilitat genètica, fonamental en els programes de millora, concretament per a l'adaptació a la producció ecològica. En la present tesi s'ha estudiat la qualitat nutricional i organolèptica del cultiu del pebrot (Capsicum sp.) en els dos estats de maduresa comercial i sota diferents pràctiques culturals (sistemes ecològic i convencional). S'han analitzat els principals compostos nutricionals dels fruits de pebrot: l'àcid ascòrbic, els compostos fenòlics i els carotenoides; així com la fracció volàtil, i s'ha estudiat la seva variació en una àmplia col·lecció varietal. Addicionalment, s'ha estudiat la fertilitat biològica de terra a través de les activitats enzimàtiques de la fosfatasa alcalina i la catalasa en interacció amb el cultiu del pebrot sota els sistemes de cultiu convencional i ecològic. L'estudi de la qualitat nutricional mostrar amplis rangs de variació segons el factor genotip, encara que l'estat de maduració i el sistema de cultiu també van contribuir a les diferències observades. En general, els fruits madurs van presentar nivells superiors de compostos bioactius, especialment per a l'àcid ascòrbic. Així mateix, el cultiu ecològic va proporcionar, en general, nivells més alts d'àcid ascòrbic i compostos fenòlics en estat madur. La notable interacció genotip × sistema de cultiu per al contingut en àcid ascòrbic i fenols totals, i en menor intensitat per al contingut en flavonoides individuals, va permetre la selecció de diverses accessions amb alt rendiment en aquests compostos, sota condicions de cultiu ecològic en els dos estats de maduració. Per contra, els efectes de el sistema de cultiu i la interacció genotip × sistema de cultiu no van resultar significatius en els nivells de carotenoides, la variació es va deure principalment al factor genotip. L'estudi de la composició volàtil va permetre trobar àmplies diferències quantitatives i qualitatives per aquest factor de qualitat organolèptica en els fruits de pebrot, depenent principalment del factor genètic i de l'estat de maduració, mentre que el sistema de cultiu va afectar en menor mesura i significativament a alguns compostos volàtils individuals. Es van trobar un total de 124 i 122 volàtils en estat immadur i madur, respectivament. Els canvis en el perfil volàtil amb la maduració van ser evidents i molt diversos segons el genotip. En general, el sistema de cultiu ecològic va afavorir l'acumulació de compostos volàtils en els dos estats de maduració i durant el procés de maduració. Les activitats enzimàtiques fosfatasa alcalina i catalasa van augmentar durant la fase de cultiu, amb activitats màximes generals en la fase intermèdia, especialment per a la fosfatasa alcalina. El sistema de cultiu va tenir una major contribució a la variació, resultat significatiu i superior per al sistema convencional en fase intermèdia per a ambdues activitats i estant generalment seguit del factor genotip, significatiu per a la fosfatasa alcalina en fase intermèdia. La interacció genotip × sistema de cultiu permetria seleccionar varietats, amb capacitat perquè els sòls acumulessin grans valors de les activitats enzimàtiques fosfatasa alcalina i catalasa, en el sistema de cultiu ecològic. En definitiva, els resultats d'aquest treball proporcionaran informació d'especial utilitat per a programes d'investigació en hortalisses d'alt valor afegit i programes de millora de la qualitat en agricultura ecològica, així com per al control d'autenticitat de varietats tradicionals. / [EN] The current environmental problem is demanding more sustainable production systems, such as organic farming. A good alternative for these systems are the traditional varieties of pepper, with characteristic quality traits and wide genetic variability, essential in breeding programs, specifically for adaptation to organic production. In this thesis, the nutritional and organoleptic quality of the pepper crop (Capsicum sp.) at the two ripening stages and under different cultural practices (ecological and conventional systems) has been studied. The main nutritional compounds of pepper fruits have been analysed: ascorbic acid, phenolic compounds and carotenoids; as well as the volatile fraction, and its variation has been studied in a wide varietal collection. Additionally, the biological fertility of the soil has been studied through the enzymatic activities of alkaline phosphatase and catalase in interaction with pepper cultivation under conventional and ecological growing systems. The study of nutritional quality showed wide ranges of variation according to the genotype factor, although the ripening stage and the growing system also contributed to the differences observed. In general, fully ripe fruits presented higher levels of bioactive compounds, especially for ascorbic acid. Also, organic farming provided, in general, higher levels of ascorbic acid and phenolic compounds at the fully ripe stage. The remarkable genotype × growing system interaction for the content of ascorbic acid and total phenolics, and to a lesser extent for the content of individual flavonoids, allowed the selection of several accessions with high yield in these compounds, under organic farming conditions at both ripening stages. On the contrary, the effects of the growing system and the genotype × growing system interaction were not significant in carotenoid levels, whose variation was mainly due to the genotype factor. The study of the volatile composition allowed to find wide quantitative and qualitative differences for this organoleptic quality factor in pepper fruits, depending mainly on the genetic factor and the ripening stage, while the growing system affected to a lesser extent and significantly some individual volatile compounds. A total of 124 and 122 volatiles were found at unripe and fully ripe stages, respectively. The changes in the volatile profile with ripening were evident and very diverse depending on the genotype. In general, the organic growing system favoured the accumulation of volatile compounds at both ripening stages and during the ripening process. Alkaline phosphatase and catalase enzymatic activities increased during the growing phase, with overall peak activities in the intermediate phase, especially for alkaline phosphatase. The growing system had a greater contribution to variation, a significant and superior result for the conventional system in the intermediate phase for both activities and being generally followed by the genotype factor, significant for alkaline phosphatase in the intermediate phase. The genotype × growing system interaction would allow the selection of varieties, with the capacity for soils to accumulate higher values of the enzymatic activities alkaline phosphatase and catalase, in the organic growing system. In conclusion, the results of this work will provide information of special use for research programs in vegetables with high added value and quality breeding programs in organic farming, as well as for the authenticity control of traditional varieties. / Authors thank seeds providers included in Table 1, like P.W. Bosland, S. Lanteri, François Jourdan, I. Ruiz de Galarreta and the different Regulatory Boards of the PDOs and GPIs included in this study. Authors also thank LA UNIÓ for the availability of experimental fields and the technical support from Manuel Figueroa, Rafael Hurtado, Antonio Muñoz and Ricard Ballester in the trials. Finally, the advice of Prof. Jaime Prohens in statistical procedures is also thanked. / Ribes Moya, AM. (2020). Selección y mejora de ecotipos de pimiento (Capsicum sp.) para agricultura sostenible y calidad nutricional y organoléptica [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/159886 / TESIS

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