Orientador: Vanderlan da Silva Bolzani / Banca: Jairo Kenupp Bastos / Banca: Patrícia Sartorelli / Resumo: O presente trabalho teve como objetivo o estudo químico da fração diclorometânica das folhas de Senna multijuga, visando o isolamento, purificação e elucidação estrutural de alcalóides piridínicos com potencial farmacológico. Inicialmente foi preparado o extrato hidroalcóolico das folhas, o qual foi submetido à partição líquido/líquido com hexano, diclorometano e acetato de etila. As quatro frações obtidas foram avaliadas por CCD, sendo a fração diclorometânica a de maior concentração alcaloídica. Essa fração foi submetida à extração ácido/base e a fração alcaloídica submetida à CCD preparativa de sílica, que foi desenvolvida com uma mistura dos solventes hexano:CHCl3:AcOEt (1,5:2:6,5), de onde foram isolados cinco alcalóides que apresentaram absorção na região do ultravioleta a 254 e 286 nm. Desta separação obteve-se dois alcalóides puros, 7'-multijuguinona (5), 12'-hidroxi-7'-multijuguinona (2), e os demais foram purificados por cromatografia líquida de alta eficiência (CLAE): 4'-multijuguinato de metila (3), 7'- multijuguinol (4a e 4b) e 12'-hidroxi-7'-multijuguinol (1a e 1b). Estas substâncias tiveram suas estruturas elucidadas pelo uso dos experimentos de Ressonância Magnética Nuclear (RMN) e Espectrometria de Massas com ionização por eletrospray (EM). A partir destas análises foi possível determinar a estrutura de sete substâncias, sendo todas inéditas na literatura. As substâncias apresentaram atividade anticolinesterásica moderada, o que justifica a importância de estudos com metabólitos secundários. Os pares 1a e 1b e 4a e 4b isolados apresentam isomeria que ainda não determinada. Pelas análises do perfil alcaloídico determinado no estudo sobre a espécie, pode-se constatar que os demais órgãos também acumulam alcalóides, porém, em concentrações diferentes, prevalencendo um ou dois deles. / Abstract: This present work deals with the chemical study of the dichloromethane fraction of the Senna multijuga leaves, aiming the isolation, purification and structural elucidation of the new bioactive pyridine alkaloids, which can be useful for further pharmacological evaluation. The leaves ethanol extract was subjected a liquid/liquid partitions with hexane, dichloromethane and ethyl acetate, resulting in four fractions, which were evaluated for TLC, being the dichloromethane the fraction with the highest concentration of alkaloids. This fraction was subjected to acid/basic extraction, and the alkaloidal fraction (1.42 g) was subjected to silica preparative TLC, and eluted with a solvent system: n-hexane:CHCl3:AcOEt (1.5:6.5:2), which were isolated five alkaloids. From this procedures were obtained alkaloids, 7'-multijuguinone (5), 12'-hydroxyl-7'- multijuguinone (2) and the resulting mixture was further purified by HPLC yielding methyl 4'-multijuguinate (3), 7'-multijuguinol (4a e 4b) and 12'-hydroxyl-7'- multijuguinol (1a e 1b). The structure of all alkaloids were determined by RMN and mass spectral data analysis resulting in seven new moderate acetylcholinesterase inhibitor derivatives. The alkaloids pairs, 1a;1b and 4a;4b, are isomeric mixtures, not identified yet. Through the alkaloid profile has been established after several data accumulated from this plant, it was observed that others organs also accumulate the same alkaloids, but in different concentration prevailing one or two alkaloids. / Mestre

Submitted by JULIA DIETSCHE MONFARDINI null (juliadm@iq.unesp.br) on 2016-04-01T16:52:10Z No. of bitstreams: 1 Dissertação_Julia Dietsche Monfardini.pdf: 10246419 bytes, checksum: dff6716d7f1e0f36c408ba7afeff1e25 (MD5) / Approved for entry into archive by Ana Paula Grisoto (grisotoana@reitoria.unesp.br) on 2016-04-05T18:12:36Z (GMT) No. of bitstreams: 1 monfardini_jd_me_araiq_par.pdf: 807325 bytes, checksum: 9a9635608b6709f23a60f0fd6f5e676e (MD5) / Made available in DSpace on 2016-04-05T18:12:36Z (GMT). No. of bitstreams: 1 monfardini_jd_me_araiq_par.pdf: 807325 bytes, checksum: 9a9635608b6709f23a60f0fd6f5e676e (MD5) Previous issue date: 2016-03-10 / Os micro-organismos têm demonstrado serem valiosas fontes de princípios ativos de uso clínico, no qual a penicilina é o exemplo mais conhecido de metabólito secundário, de importância medicinal, produzido por fungos. A espécie vegetal Eugenia jambolana Lam., popularmente conhecida como Jambolão, é utilizada na medicina popular brasileira, principalmente no tratamento de diabetes. O fungo Botryosphaeria parva, isolado das folhas da E. jambolana (L.) e codificado como Ej_F01, foi cultivado em escala ampliada em três meios de cultivo: Czapek®, PDB e Milho. Após o período de fermentação do endófito no meio de milho, foi feita uma extração direta com MeOH, seguida de filtração e evaporação do solvente. O extrato obtido foi solubilizado em AcOEt e fez-se três partições líquido-líquido com H2O, após a evaporação do solvente orgânico, o extrato foi dissolvido em CH3CN e foi feita uma nova partição líquido-líquido com Hexano, após a evaporação da CH3CN, foi obtido o extrato de interesse. Após o período de fermentação do endófito nos meios líquidos (PDB e Czapek®), a suspensão micelar foi filtrada a pressão reduzida, separando os micélios dos meios fermentados. Em seguida, os filtrados aquosos foram submetidos a três partições líquido-líquido com AcOEt, e após a evaporação do solvente orgânico, foram obtidos os extratos brutos de interesse. Todos os extratos foram fracionados utilizando técnicas cromatográficas, como Cromatografia em Coluna e/ou Cromatografia Líquida de Alta Eficiência preparativa. Após o fracionamento dos extratos brutos, foi possível obter sete substâncias oriundas do extrato do milho, seis substâncias do extrato do Czapek®, e uma do extrato de PDB, que foram submetidas a análises espectrométricas (EM, RMN de 1H, 13C, 1D e 2D) para a determinação/identificação estrutural. As substâncias 05, 08, 09 e 11 foram identificadas como 5-hidroximeleina, rel. (3S, 4S)-4-hidroximeleina, meleina e rel. (3S, 4R)-4-hidroximeleina, respectivamente, pertencentes à classe das isocumarinas; as substâncias 01, 03, 06 e 07 foram identificadas como botryosphaerin A, 13,14,15,16-tetranorlabd-7-eno-19,6β:12,17-diolídeo, CJ-14445 e oidiolactona E, respectivamente, pertencentes à classe dos diterpenos tetranorlabdanos; a substância 12 foi identificada como botryosphaerona D, pertencente à classe das naftalenonas. A substância 02, inédita, foi nomeada como rel. (4S, 5R, 6R, 10S)-18-hidroxi-13,14,15,16-tetranorlabd-7,9-dieno-19,6β:12,17-diolídeo, codificada como botryosphaerin I. Os estudos relacionados ao B. parva têm como finalidade verificar sua produção metabólica em diferentes meios de cultivo, bem como a avaliação do potencial biológico dos extratos brutos, frente aos ensaios antioxidante, antifúngico e anticolinesterásico. / Microorganisms have proven to be valuable sources of active principles for clinical use, wherein the penicillin is the best known example of a secondary metabolite of medicinal importance, produced by fungi. The plant species Eugenia jambolana Lam., commonly called Black Plum, is used in traditional Brazilian medicine, especially in the treatment of diabetes. The Botryosphaeria parva fungus, isolated from leaves of E. jambolana (L.) encoded as Ej_F01, was grown on an enlarged scale in three different culture media: Czapek®, PDB and corn. After the fermentation period in the corn media, a direct extraction was done with MeOH followed by filtration and evaporation of the solvent. The extract was disolved in AcOEt and made three liquid-liquid partition with H2O, after evaporation of the organic solvent, the extract was dissolved in CH3CN and was made a new liquid-liquid partition with Hexane, after evaporation of the CH3CN, it was obtained extract of interest. After the fermentation period in liquid media (PDB and Czapek®), the micellar suspension was filtered under reduced pressure, separating the mycelium from the fermented broth. Then, the aqueous filtrates were subjected to three liquid-liquid partition with AcOEt, and after evaporation of the organic solvent, there were obtained the crude extracts of interest. All extracts were fractionated using chromatographic techniques such as column chromatography and/or preparative High Performance Liquid Chromatography efficiency. After the fractioning of crude extracts, it was possible to obtain seven substances derived from corn extract, six substances obtained from Czapek extract, and a substance obtained from the PDB extract, which were submitted to spectrometric analysis (MS-ESI, NMR 1H, 13C, 1D and 2D) for determining and structural identification. The substances 05, 08, 09 and 11 were identified as 5-hydroxymelein, rel. (3S, 4S)-4-hydroxymelein, Melein and rel. (3S, 4R)-4-hydroxymelein, respectively, belonging to the class of isocoumarins; the substances 01, 03, 06 and 07 were identified as botryosphaerin A, 13,14,15,16-tetranorlabd-7-ene-19,6β:12,17-diolide, CJ-14445 and oidiolactone E, respectively, belonging to the class of tetranorlabdanes diterpenoids, and substance 12 was identified as botryosphaerone D, belonging to the class of naftalenones. A new tetranorlabdane diterpenoid was obtained, substance 02, named rel. (4S, 5R, 6R,10S)-18-hydroxy-13,14,15,16-tetranorlabd-7,9-dien-19,6β:12,17-diolide and encoded botryosphaerin I. These studies related to the B. parva are intended to verify your metabolic production in different culture media as well as the evaluation of the biological potential of extracts compared to antioxidant, antifungal and anticholinesterase trials.

Made available in DSpace on 2014-06-11T19:29:11Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-02-16Bitstream added on 2014-06-13T20:19:05Z : No. of bitstreams: 1 francisco_w_me_araiq.pdf: 5726334 bytes, checksum: fad4f57aaf53ae933a5c86aa96974115 (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O presente trabalho teve como objetivo o estudo químico da fração diclorometânica das folhas de Senna multijuga, visando o isolamento, purificação e elucidação estrutural de alcalóides piridínicos com potencial farmacológico. Inicialmente foi preparado o extrato hidroalcóolico das folhas, o qual foi submetido à partição líquido/líquido com hexano, diclorometano e acetato de etila. As quatro frações obtidas foram avaliadas por CCD, sendo a fração diclorometânica a de maior concentração alcaloídica. Essa fração foi submetida à extração ácido/base e a fração alcaloídica submetida à CCD preparativa de sílica, que foi desenvolvida com uma mistura dos solventes hexano:CHCl3:AcOEt (1,5:2:6,5), de onde foram isolados cinco alcalóides que apresentaram absorção na região do ultravioleta a 254 e 286 nm. Desta separação obteve-se dois alcalóides puros, 7'-multijuguinona (5), 12'-hidroxi-7'-multijuguinona (2), e os demais foram purificados por cromatografia líquida de alta eficiência (CLAE): 4'-multijuguinato de metila (3), 7'- multijuguinol (4a e 4b) e 12'-hidroxi-7'-multijuguinol (1a e 1b). Estas substâncias tiveram suas estruturas elucidadas pelo uso dos experimentos de Ressonância Magnética Nuclear (RMN) e Espectrometria de Massas com ionização por eletrospray (EM). A partir destas análises foi possível determinar a estrutura de sete substâncias, sendo todas inéditas na literatura. As substâncias apresentaram atividade anticolinesterásica moderada, o que justifica a importância de estudos com metabólitos secundários. Os pares 1a e 1b e 4a e 4b isolados apresentam isomeria que ainda não determinada. Pelas análises do perfil alcaloídico determinado no estudo sobre a espécie, pode-se constatar que os demais órgãos também acumulam alcalóides, porém, em concentrações diferentes, prevalencendo um ou dois deles. / This present work deals with the chemical study of the dichloromethane fraction of the Senna multijuga leaves, aiming the isolation, purification and structural elucidation of the new bioactive pyridine alkaloids, which can be useful for further pharmacological evaluation. The leaves ethanol extract was subjected a liquid/liquid partitions with hexane, dichloromethane and ethyl acetate, resulting in four fractions, which were evaluated for TLC, being the dichloromethane the fraction with the highest concentration of alkaloids. This fraction was subjected to acid/basic extraction, and the alkaloidal fraction (1.42 g) was subjected to silica preparative TLC, and eluted with a solvent system: n-hexane:CHCl3:AcOEt (1.5:6.5:2), which were isolated five alkaloids. From this procedures were obtained alkaloids, 7'-multijuguinone (5), 12'-hydroxyl-7'- multijuguinone (2) and the resulting mixture was further purified by HPLC yielding methyl 4'-multijuguinate (3), 7'-multijuguinol (4a e 4b) and 12'-hydroxyl-7'- multijuguinol (1a e 1b). The structure of all alkaloids were determined by RMN and mass spectral data analysis resulting in seven new moderate acetylcholinesterase inhibitor derivatives. The alkaloids pairs, 1a;1b and 4a;4b, are isomeric mixtures, not identified yet. Through the alkaloid profile has been established after several data accumulated from this plant, it was observed that others organs also accumulate the same alkaloids, but in different concentration prevailing one or two alkaloids.

L'objectif de la présente étude est l’évaluation des effets de l'exposition gestationnelle et lactationnelle aux anguilles contaminées aux POP dont la Σ6 PCB-NDL, sur la mémoire, la capacité d’apprentissage, l’activité motrice, le comportement de résignation, le statut oxydatif et l'activité de l'acétylcholinestérase des souris de génération F1. Pour ce faire, des souris femelles ont été exposées, à partir du sixième jour de gestation (JG 6) jusqu'au jour postnatal 21 (JPN 21), par voie d'ingestion, aux anguilles contaminées contenant Σ 6 PCB-NDL. Trois doses de la Σ 6 PCB-NDL ont été choisies : 85, 216 et 400ng/kg/j en se référant à un groupe de souris témoins. Nos résultats ont montré que l'exposition gestationnelle et lactationnelle, aux anguilles contaminées, contenant la Σ 6 PCB-NDL, n’a pas affecté la capacité d’apprentissage, ni la mémoire. Cependant, le traitement, utilisé dans cette étude, pourrait causer une hyperactivité tardive, persistante et sexe dépendante qui ne touche les souris femelles de la génération F1. En outre, un effet « depresive-like » de l’exposition gestationnelle et lactationnelle des souris de génération F1 aux anguilles contaminée par les POP y inclus la Σ 6 PCB-NDL, a été relevé de façon sexe dépendante. Le comportement « depresive-like » ne touche que les souris mâles et il était persistant au fil de temps. L’étude du statut oxydatif des trois régions cérébrales (cortex préfrontal, hippocampe et cervelet), a révélé que le traitement pourrait perturber, de façon sexe dépendante, l’activité des enzymes antioxydants (Cat, SOD) au niveau du cortex préfrontal et de l’hippocampe. Les conséquences du stress oxydant, dans cette étude se sont manifestées par une augmentation du taux des protéines carbonylées au niveau du cortex préfrontal et de l’hippocampe de certains groupes de souris mâles traitées. Cependant, aucune différence significative, n’a été enregistrée, au niveau des trois régions cérébrales étudiées, dans le taux de MDA, entre les souris traitées et les souris témoins. La perturbation du statut oxydatif a été sexe dépendante, elle n’a touché que les souris mâles. L’étude de l’activité de l’acétylcholinestérase a montré que l’ingestion des souris gestantes et puis allaitantes des anguilles contaminées contenant Σ6PCB-NDL aux doses 85 et 400 ng / kg / jour était à l’origine d’une inhibition significative, par rapport au témoin de l’activité de l’acétylcholinestérase, au niveau du cortex préfrontal de leurs progéniture femelles. Alors qu’au niveau de l’hippocampe, une inhibition significative de l’activité de cette enzyme a été enregistrée chez les souris mâles traitées aux anguilles contenant Σ6PCB-NDL à la dose de 85 ng / kg / jour et chez toute la progéniture de souris femelles des groupes traitées / The objective of the present study is the evaluation of gestational and lactational exposure to contaminated eels, containing Σ6 NDL-PCBs, on learning, memory, motor activity, resignation behavior, oxidative status and acetylcholinesterase activity of the offspring mice. For this purpose female mice were orally exposed during gestational and breast feeding period (from JG6 to JPN 21), through standard diet containing contaminated eel with Σ6 NDL-PCBs or no. Three doses of Σ 6 NDL-PCBs were chosen: 85ng / kg / day, 216 ng / kg / day and 400ng / kg / day. A control group of female mice, receiving the standard diet without fish, was used. Results showed that gestational and lactational exposure to contaminated eels, containing Σ6 NDL-PCBs, did not affect the memory and learning capacities. However the early-life exposure to polluted eels containing Σ6 NDL-PCBs caused late, persistent and gender dependent neurobehavioral hyperactive effects in offspring mice that affects only offspring females. Furthermore our results suggest that the consumption of polluted fish by pregnant and lactating dams might cause neurobehavioral depression only in their male offspring, at progressive life stage. Moreover the study of oxidative status on the three brain areas (prefrontal cortex, hippocampus and cerebellum), revealed that the treatment had affected the activity of antioxidant enzymes (Cat, SOD) only in the prefrontal cortex, hippocampus of male mice. The consequences of oxidative stress in this study were revealed by an increase of protein carbonyl levels in the hippocampus and the prefrontal cortex of offspring male mice, exposed to chemically contaminated fish containing Σ6 NDL-PCBs at the dose of 85 and 216 ng / kg / day, respectively. However the treatment had no effect on the lipids. Indeed there were no significant differences in the MDA rate, in the three studied regions of the brain, between treated and control mice. Moreover, gestational and lactational exposure to contaminated eels, containing Σ6NDL-PCB at the dose of 85 and 400 ng / kg / day, had inhibited acetylcholinesterase activity, in the prefrontal cortex, only of female mice. However, in the hippocampus, the enzyme activity was inhibited of male mice, exposed to contaminated eels, containing Σ6NDL-PCB at the dose of 85 ng/ kg / day and in all the treated female offspring mice

<p>&nbsp;&nbsp; Cilj ove doktorske disertacije predstavljao je ispitivanje fitohemijskog sastava i biolo&scaron;ke aktivnosti vodenih i metanolnih ekstrakata svežih i suvih plodova, kao&nbsp; i pekmeza&nbsp; pripremljenog&nbsp; po tradicionalnoj recepturi od plodova&nbsp; četiri (od kojih se jedna javlja u dve forme)&nbsp; samonike&nbsp; vrsta&nbsp; roda&nbsp; <em>Sorbus</em>&nbsp; L.:&nbsp; S.<em> aucuparia</em>, S.<em> domestica</em>, S. <em>torminalis</em>&nbsp; f.&nbsp; t<em>orminalis</em>, S. <em>torminalis</em>&nbsp; f. <em>semitorminalis</em>&nbsp; i&nbsp; S. intermedia Ispitivanje fitohemijskog sastava obuhvatalo je LC-MS/MS analizu&nbsp; 44 odabrana fenolna jedinjenja&nbsp; i&nbsp; hinske kiseline (organska&nbsp; kiselina). Takođe, spektrofotometrijski je&nbsp; određen&nbsp; sadržaj&nbsp; ukupnih fenolnih i flavonoidnih jedinjenja, kao i&nbsp; sadržaj askorbinske kiseline. Evaluacija biolo&scaron;ke aktivnosti obuhvatala je&nbsp; <em>in vitro&nbsp; </em>ispitivanja antioksidantne aktivnosti, kao i ispitivanje uticaja ekstrakata odabranih vrsta&nbsp; roda&nbsp;<em> Sorbus</em>&nbsp; na aktivnost enzima acetilholinesteraze, antimikrobni, kao i antiproliferativni potencijal.</p><p>&nbsp;&nbsp; Sumiranjem dobijenih rezultata može se zaključiti da sveži i suvi plodovi ispitivanih vrsta&nbsp; <em>Sorbus</em>,&nbsp; kao i&nbsp; pekmezi&nbsp; predstavljaju&nbsp; umeren izvor&nbsp; fenolnih jedinjenja.&nbsp; Kao najzastupljenije fenolne kiseline izdvojile su se protokatehinska i&nbsp; ferulna, a među flavonoidima amentoflavon i kvercetin-3<em>-O-</em>glukozid.&nbsp; Pored toga, hinska kiselina je zabeležena u značajnoj količini u svim analiziranim ekstraktima.</p><p>&nbsp;&nbsp;&nbsp; Ekstrakti ispitivanih vrsta pokazali su&nbsp; umeren&nbsp; antioksidantni potencijal koji se ogleda u njihovoj sposobnosti neutralizacije nekoliko radikalskih vrsta, redukcionom potencijalu i sposobnosti inhibicije lipidne peroksidacije.&nbsp; Ispitivanjem uticaja ekstrakata odabranih vrsta&nbsp;<em> Sorbus</em>&nbsp; naaktivnost&nbsp; enzima&nbsp; acetilholinesteraze&nbsp; dokazana&nbsp; je&nbsp; jedino&nbsp; umerena aktivnost&nbsp; ekstrakata vrste&nbsp; <em>S. aucuparia</em>.&nbsp; Takođe, ispitivani ekstrakti&nbsp; vrsta roda&nbsp; <em>Sorbus</em>&nbsp; ispoljili su umerenu&nbsp; antimikrobnu&nbsp; aktivnost u pogledu inhibicije&nbsp; rasta Gram pozitivne bakterije,&nbsp; <em>Staphylococcus aureus</em>, i Gram negativne bakterije,&nbsp;<em> Escherichia coli</em>.&nbsp; Vodeni i metanolni ekstrakti svežih i suvih plodova vrste&nbsp; <em>S. aucuparia</em>&nbsp; pokazali su umeren i inhibitorni potencijal prema rastu tumorskih (HeLa, MCF7, HT-29), ali i netumorskih ćelijskih linija (MRC-5).&nbsp; Rezultati dobijeni u ovoj doktorskoj distertaciji&nbsp; ukazuju na značajan&nbsp; biopotencijal plodova i pekmeza&nbsp; ispitivanih vrsta&nbsp;<em> Sorbus</em>&nbsp; i ukazuju na njihovu primenu u prehrambenoj industriji u vidu funkcionalne hrane.</p> / <p>The aim of presented&nbsp; PhD&nbsp; thesis was investigation of phytochemical composition and biological activity of water and methanol extracts of fresh and air-dried&nbsp; fruits, as&nbsp;&nbsp; well&nbsp; as&nbsp; jam,&nbsp; made according to traditional recipe,&nbsp; of fruits&nbsp; of four&nbsp; (one of them occurs in two forms)&nbsp; wild growing&nbsp; <em>Sorbus</em>&nbsp; L. species:&nbsp; <em>S. aucuparia, S. domestica, S. torminalis&nbsp; f. torminalis, S. torminalis f. semitorminalis</em> and <em>S. intermedia</em>. Examination of phytochemical composition included LC-MS/MS analysis of 44 selected phenolic compounds&nbsp; and&nbsp; quinic acid (organic acid).&nbsp; Also, total phenolic and flavonoid contents, as well as&nbsp; ascorbic acid&nbsp; content,&nbsp; were determined spectrophotometrically. Biological activity evaluation of extracts of<em>&nbsp; Sorbus</em>&nbsp; species included&nbsp;<em> in vitro&nbsp;</em> investigation of antioxidant, anti-acetylcholinesterase, antimicrobial&nbsp; and cytotoxic activity.</p><p>&nbsp;&nbsp; According to obtained results, fresh and air-dried&nbsp; fruits, as well as jam&nbsp; present&nbsp; moderate&nbsp; source of phenolic compounds. Amongs examined phenolic acids protocatechuic&nbsp; and ferulic acids were the most abundant, and amongs investigated flavonoids amentoflavone and quercetin-3-<em>O</em>-glucoside wete present in noticeable amount.&nbsp; Furthermore, high concentration of quinic acid was present in all examined extracts.</p><p>&nbsp;&nbsp; Extracts of all examined species showed&nbsp; moderate&nbsp;&nbsp; antioxidant activity in terms of radical scavenging ability, reduction potential and inhibition of lipid peroxidation.&nbsp; Also, investigation of&nbsp; anti-acetylcholinesterase activity revealed moderate activity only of extracts of <em>S. aucuparia</em>. Furthermore, examinated extracts of<em> Sorbus</em> species showedmoderate antimicrobial activity against Gram&ndash;positive bacteria, <em>Staphylococcus aureus</em>, and Gram-negative bacteria,&nbsp;<em> Escherichia coli</em>.&nbsp; In addition, water&nbsp; and methanol&nbsp; extracts of fresh and air-dried&nbsp; fruits of&nbsp;<em> S.aucuparia</em>&nbsp; showed&nbsp; inhibitory activity toward&nbsp; tumor (HeLa, MCF7, HT-29), and also non-tumor (MRC-5) cell lines. Presented results indicate significant&nbsp; biopotential of examined&nbsp; fruits of&nbsp; <em>Sorbus&nbsp;</em> species&nbsp; and&nbsp; support their use in food industry as functional food.</p>

<p>Prema postavljenim ciljevima u ovoj doktorskoj disertaciji sakupljeno je i&nbsp;<br />determinisano&nbsp;ukupno&nbsp;sedam&nbsp;vrsta&nbsp;autohtonih&nbsp;gljiva&nbsp;sa&nbsp;područja&nbsp;Fru&scaron;ke&nbsp;gore, Tare i&nbsp; Vr&scaron;ačkog brega, pet lignikolnih&nbsp;‐&nbsp;<em>Bjerkandera adusta</em>,&nbsp;<em>Pleurocybella&nbsp; porrigens</em>,&nbsp;St<em>ereum&nbsp;hirsutum,&nbsp;Stereum&nbsp;subtomentosum</em>&nbsp;i&nbsp;Trametes&nbsp;versicolor,&nbsp;i&nbsp;<br />dve&nbsp; terikolne&nbsp;‐&nbsp;<em>Amanita&nbsp; strobiliformis</em>&nbsp;i Hydnum repandum. Utvrđena je&nbsp; njihova&nbsp;biolo&scaron;ka&nbsp;aktivnost&nbsp;(antiradikalska,&nbsp;antioksidativna,&nbsp;antimikrobna,&nbsp;anti‐acetilholinesterazna&nbsp;i&nbsp;citotokisčna)&nbsp;spram&nbsp;hemijskog&nbsp;sastava&nbsp;njihovih&nbsp;vodenih&nbsp;<br />(H₂O),&nbsp;etanolnih&nbsp;(EtOH),&nbsp;metanolnih&nbsp;(MeOH)&nbsp;i&nbsp;polisaharidnih&nbsp;(PSH)&nbsp;ekstrakata.&nbsp;<br />Analiza&nbsp;hemijskog&nbsp;sastava&nbsp;odabranih&nbsp;vrsta&nbsp;uključila&nbsp;je&nbsp;određivanje&nbsp;hemijske&nbsp;<br />karakterizacije&nbsp;PSH&nbsp;ekstrakata&nbsp;‐&nbsp;FTIR&nbsp;analizom,&nbsp;određivanje&nbsp;fenolnog&nbsp;profila&nbsp;‐&nbsp;<br />HPMC/MS‐MS,&nbsp;sadržaja&nbsp;organskih&nbsp;kiselina&nbsp;‐&nbsp;HPLC,&nbsp;sadržaja&nbsp;masnih&nbsp;kiselina&nbsp;‐&nbsp;<br />GC‐FID&nbsp;i&nbsp;sadržaja&nbsp;biogenih&nbsp;elemenata&nbsp;‐&nbsp;AAS.&nbsp;Spektrofotometrijskim&nbsp;metodama<br />određen je ukupan sadržaj proteina i ukupan sadržaj fenola i flavonoida.&nbsp;<br />Prema&nbsp;antiradikalskoj&nbsp;aktivnosti&nbsp;OH^{&bull; ,&nbsp;}O2^&bull;‐,&nbsp;OH^&bull;,&nbsp;Asc^&bull;,&nbsp;DPPH^&bull;&nbsp; i&nbsp;ABTS^&bull;+&nbsp;izdvojili&nbsp;<br />su&nbsp; se&nbsp; ekstrakti lignikolnih vrsta:&nbsp; MeOH ekstrakt vrste&nbsp;<em>P. porrigens</em>, H₂O&nbsp; ekstrakt&nbsp;<em>P.&nbsp;porrigens</em>,&nbsp;MeOH&nbsp;ekstrakt&nbsp;<em>T.&nbsp;versicolor</em>,&nbsp;H₂O&nbsp;ekstrakt&nbsp;<em>S.&nbsp;hirsutum, </em>MeOH&nbsp;ekstrakt&nbsp;<em>S.&nbsp;subtomentosum</em>&nbsp;i&nbsp;H₂O&nbsp;ekstrakt&nbsp;<em>B.&nbsp;adusta</em>,&nbsp;navedenim&nbsp;redom.&nbsp;<br />Najjaču antioksidativnu aktivnost dobijenu FRAP i polarografskom HPMC&nbsp;<br />metodom ispoljili su PSH i H₂O ekstrakti terikolne vrste&nbsp;<em>A. strobiliformis</em>.&nbsp;<br />Antimikrobna aktivnost analiziranih ekstrakata određena je ispitivanjem&nbsp;<br />antibakterijskog,&nbsp;antifungalnog&nbsp;i&nbsp;antiviralnog&nbsp;potencijala,&nbsp;pri&nbsp;čemu&nbsp;se&nbsp;izdvojila&nbsp;<br />vrsta&nbsp;<em> H.&nbsp; repandum</em>&nbsp;ispoljavajući najbolji efekat na Gram‐pozitivne i Gram‐<br />negativne bakterije i na sve analizirane fitopatogene izolate&nbsp;<em>(Fusarium&nbsp;</em>i&nbsp;<br />Alternaria)&nbsp;i<em>&nbsp;T.&nbsp;versicolor</em>&nbsp;na&nbsp;analizirani&nbsp;bakteriofag.&nbsp;Anti‐acetilholinesterazna&nbsp;<br />aktivnost određena je testovima in solid i in liquid, a najbolji procenat&nbsp;<br />inhibicije AChE ispoljili su EtOH ekstrakti vrsta&nbsp;<em>S. hirsutum</em>,&nbsp;<em>B. adusta</em>,&nbsp;<em>S</em>.&nbsp;<br /><em>subtomentosum</em>&nbsp;i&nbsp;<em>T.&nbsp;versicolor</em>.&nbsp;Citotoksična&nbsp;aktivnost&nbsp;ekstrakata&nbsp;određena&nbsp;je&nbsp;<br />MTT testom, a prema najboljoj ispoljenoj aktivnosti izdvojili su se MeOH&nbsp;<br />ekstrakt<em>&nbsp;P.&nbsp;porrigens&nbsp;</em>i&nbsp;ekstrakti<em>&nbsp;B.&nbsp;adusta</em>,&nbsp;H₂O&nbsp;i&nbsp;EtOH.&nbsp;Citotoksična&nbsp;aktivnost&nbsp;<br />ovih&nbsp;lignikolnih&nbsp;vrsta&nbsp;naročito&nbsp;je&nbsp;izražena&nbsp;nakon&nbsp;72&nbsp;h.&nbsp;Na&nbsp;osnovu&nbsp;dobijenih&nbsp;<br />rezultata, gde su se istakle različite vrste i njihovi različiti ekstrakti u&nbsp;<br />primenjenim&nbsp;testovima,&nbsp;jasno&nbsp;je&nbsp;da&nbsp;biolo&scaron;ka&nbsp;aktivnost&nbsp;i&nbsp;hemijski&nbsp;sastav&nbsp;zavise&nbsp;<br />od&nbsp;porekla,&nbsp;vrste&nbsp;i&nbsp;tipa&nbsp;ekstrakta&nbsp;analiziranih&nbsp;gljiva.&nbsp;Na&nbsp;osnovu&nbsp;tipa&nbsp;rastvarača&nbsp;<br />odnosno ekstrakata vrsta, koje su pokazale najbolju aktivnost spram pomenutih&nbsp; testova i na osnovu dobijenih korelacija kao i na osnovu detektovanih jedinjenja,&nbsp; pretpostavljamo da su za ispoljene aktivnosti u najvećoj meri odgovorna fenolna&nbsp; jedinjenja&nbsp;i&nbsp;polisaharidi.&nbsp;&nbsp;<br />&nbsp;<br />Ispoljeni&nbsp;biopotencijal&nbsp;analiziranih&nbsp;vrsta&nbsp;gljiva&nbsp;upućuje&nbsp;na&nbsp;njihovu&nbsp;potencijalnu&nbsp;<br />upotrebu kao funkcionalne hrane i nutraceutika, kao i u biokontroli&nbsp;<br />fitopatogena.</p> / <p>According&nbsp; to&nbsp; the&nbsp; set&nbsp; aims&nbsp; of&nbsp; the&nbsp; presented&nbsp; PhD&nbsp; thesis,&nbsp; seven&nbsp; autochthonous fungal species from the region of Fruska Gora, Tara&nbsp; and Vr&scaron;ac Mountains were collected and determined: five lignicolous ‐ <em>Bjerkandera&nbsp; adusta,&nbsp; Pleurocybella&nbsp; porrigens,&nbsp; Stereum&nbsp; hirsutum,&nbsp; Stereum subtomentosum and Trametes versicolor</em>, and two terricolous ‐ <em>Amanita strobiliformis </em>and <em>Hydnum repandum</em>. Biological activity of these&nbsp; species&nbsp; (antiradical,&nbsp; antioxidant,&nbsp; antimicrobial,&nbsp; anti‐ acetylcholinesterase and cytotoxic) was determined in relation to the chemical composition of the extracts, aqueous (H₂O), ethanolic (EtOH), methanolic (MeOH) and polysaccharide (PSH). Analysis of the chemical content of analyzed species included chemical characterization of PSH extracts&nbsp; &ndash;&nbsp; by&nbsp; FTIR&nbsp; analysis,&nbsp; determination&nbsp; of&nbsp; phenolic&nbsp; profile ‐ by HPMC/MS‐MS, content of organic acids ‐ by HPLC, fatty acid content ‐ by&nbsp; GC‐FID&nbsp; and&nbsp; content&nbsp; of&nbsp; biogenic&nbsp; elements ‐ by&nbsp; AAS. Spectrophotometric methods were applied for determination of the content of total proteins, polyphenols and flavonoids. According to the antiradical activity obtained towards OH^&bull;, О2^&bull;‐, OH^&bull;, Asc^&bull;, DPPH^&bull;&nbsp;and ABTS^&bull;+ extracts of lignicolous species were singled out: <em>P. porrigens&nbsp;</em> (MeOH&nbsp; extract),&nbsp;<em> P.&nbsp; porrigens</em>&nbsp; (H₂O&nbsp; extract),&nbsp;<em>T. versicolor</em>&nbsp;(MeOH&nbsp;&nbsp; extract),<em> S. hirsutum </em>(H₂O extract), <em>S. subtomentosum</em> (MeOH extract) and&nbsp;<em>B. austa</em> (H₂O&nbsp; extract),&nbsp; respectively. The&nbsp; highest antioxidant activity obtained by FRAP and the polarographic HPMC method was exhibited&nbsp; for PSH&nbsp; and&nbsp; H₂O extracts of the terricolous species&nbsp;<em>A. strobiliformis</em>. The intimicrobial activity of analyzed extracts was determined by examination of antibacterial, antifungal and antiviral potentials, whereby&nbsp; the species&nbsp; <em>H.&nbsp; repandum&nbsp;</em>was separated by exhibiting the best effect on Gram‐positive&nbsp; and Gram‐negative bacteria, and all the analyzed hytopathogenic isolates (<em>Fusarium, Alternaria</em>)&nbsp; and <em>T. versicolor</em>&nbsp;against analyzed bacteriophage. Anti‐cetylcholinesterase activity was determined by tests in solid and in liquid, while the best&nbsp; percent of AChE inhibition was showed by EtOH extracts of the species <em>S. hirsutum, B. adusta, S. subtomentosum </em>and <em>T. versicolor</em>.bThe cytotoxic activity of extracts was determined by MTT assay, and according to the best activity, the MeOH extract of <em>P. porrigens</em>, and H₂O and EtOH extracts of <em>B. adusta</em>&nbsp;were distinguished particularly after 72 h. Based on the results obtained, favoring different species and their different extracts in the applied tests, it is clear that the biological activity and chemical composition depend on the origin, species and type of extract of the analyzed fungi. Based on the type of solvent or extract of the species that showed the best activity in relation to the above tests and on the basis of the obtained correlations as well as on&nbsp; the basis of the detected compounds, we assume&nbsp; that&nbsp; the&nbsp; phenol compounds&nbsp; and&nbsp; polysaccharides&nbsp; are responsible for the activities performed.<br />Demonstrated bio‐potential of analyzed fungal species indicates their&nbsp; potential use as functional foods and nutraceutics, as well as in the biocontrol of phytopathogens.</p>

<p>&nbsp;&nbsp; Cilj ove doktorske disertacije predstavljalo je ispitivanje fitohemijskog sastava i biolo&scaron;ke aktivnosti vodenih i metanolnih ekstrakata svežih i suvih plodova, kao i voćne ka&scaron;e i džema pripremljenih po tradicionalnoj recepturi od plodova &scaron;est samoniklih vrsta <em>Rosa L.: R. canina, R. dumalis, R. dumetorum, R. tomentosa, R. arvensis, R. sempervirens. </em>Ispitivanje fitohemijskog sastava obuhvatalo je LC -MS/MS analizu 64 odabrana fenolna jedinjenja, hinske kiseline (organske kiseline) kao i tri triterpenoida. Takođe, izvr&scaron;eno je spektrofotometrijsko određivanje sadržaja ukupnih fenolnih i flavonoidnih jedinjenja, kao i vitamina C. Evaluacija biolo&scaron;ke aktivnosti obuhvatala je in vitro ispitivanja&nbsp; antioksidantne i antiinflamatorne aktivnosti, kao i ispitivanje uticaja ekstrakata odabranih vrsta Rosa na aktivnost enzima acetilholinesteraze i rast tumorskih i netumorskih ćelija.</p><p>&nbsp;&nbsp; Sumiranjem dobijenih rezultata može se zaključiti da sveži i suvi plodovi ispitivanih vrsta <em>Rosa</em>, kao i voćne ka&scaron;e i džemovi predstavljaju značajan izvor vitamina C i fenolnih jedinjenja, sa elagnom kiselinom kao najzastupljenijom fenolnom komponentom. Takođe u pojedinim ekstraktima vrsta<em> R. dumetorum</em> i<em> R. tomentosa</em> detektovana je visoka koncentracija ursolne kiseline, dok je hinska kiselina prisutna u značajnoj koncentraciji u svim ispitivanim ekstraktima.</p><p>&nbsp; &nbsp;Ekstrakti ispitivanih vrsta, izuzev vrste<em> Rosa arvensis</em>, pokazali su visok antioksidantni potencijal koji se ogleda u njihovoj sposobnosti neutralizacije nekoliko radikalskih vrsta, redukcionom potencijalu i sposobnosti inhibicije lipidne peroksidacije. Ispitivani ekstrakti ispoljili su umerenu antiinflamatornu aktivnost u pogledu inhibicije produkcije odabranih metabolita ciklooksigenaznog (12-HHT, TXB₂, PGE₂) i 12-lipooksigenaznog (12-HETE) metaboličkog puta arahidonske kiseline,&nbsp; posebno prostaglandina E&nbsp; . Takođe, in vitro ispitivanjem uticaja ekstrakata odabranih vrsta Rosa na aktivnost acetilholinesteraze pokazana je umerena aktivnost. Vodeni ekstrakti i ekstrakti voćnih ka&scaron;a vrsta <em>R. canina, R. tomentosa i R. sempervirens</em> pokazali su umereni inhibitorni potencijal prema rastu HeLa ćelijske linije, dok su ekstrakti vrste <em>R. sempervirens </em>pokazali aktivnost i prema HT-29 ćelijama. Dobijeni rezultati ukazuju na značajni potencijal plodova i tradicionalnih proizvoda ispitivanih vrsta <em>Rosa</em> za upotrebu u proizvodnji nutraceutika i funkcionalne hrane.</p> / <p>&nbsp;&nbsp; The aim of presented doctoral thesis was investigation of phytochemical composition and biological activity of water and methanol extracts of fresh and air-&nbsp; dried rose hips, as well as pur&eacute;e and jam made according to traditional recipe of&nbsp; hips of six wild growing<em> Rosa L. species: R. canina, R. dumalis, R. dumetorum, R</em>.&nbsp; <em>tomentosa, R. arvensis, and R. sempervirens.</em> Examination of phytochemical composition included LC-MS/MS analysis of 64 selected phenolic compounds, quinic&nbsp; acid (organic acid) and three triterpenoids. Also, total phenolic and flavonoid contents, as well as vitamin C contents were determined spectrophotometrically. Biological activity evaluation of extracts of six <em>Rosa</em> species included in vitro investigation of antioxidant, anti- inflammatory, anti-acetylcholinesterase and cytotoxic activity.<br />&nbsp;&nbsp; According to obtained results, fresh and air-dried rose hips, as well as their preserves present valuable source of vitamin C and phenolic compounds, especially&nbsp; ellagic acid, which was the most abundant examined phenolic compound. Furthermore, high concentration of ursolic acid was detected in some<em> Rosa &nbsp;</em> <em>tomentosa and R.&nbsp; rumetorum</em> extracts, while notable concentration of quinic acid&nbsp; was present in all examined extracts.<br />&nbsp;&nbsp; Extracts of all examined species, apart from <em>Rosa arvensis</em>, showed considerable antioxidant activity in terms of radical scavenging ability, reduction potential and inhibition of lipid peroxidation. Moreover, extracts exhibited moderate anti- inflammatory activity by means of inhibition of the main arachidonic acid metabolites &nbsp; formed incyclooxygenase-1 (12-HHT, TXB₂, PGE ₂ ) and 12-lipoxygenase (12-HETE)&nbsp; pathway, especially prostaglandin E₂ . Also, investigation of anti- cetylcholinesterase&nbsp; activity revealed moderate activity of extracts of all examined species. In addition,&nbsp; predominantly water extracts of fresh andair-dried rose hips, as well as pur&eacute;e of<em> R</em>.&nbsp; <em>canina, R. tomentosa </em>and <em>R. sempervirens</em> showed inhibitory activity toward HeLa, while <em>R.sempervirens</em> extracts also inhibited HT-29 cell growth. Presented results indicate significant potential of examined rose hips and their preserves for use as nutraceuticals and functional food.</p>