<p>U ovom radu su ispitani hemijski sastav i biolo&scaron;ka aktivnost vrsta&nbsp;<em> Brassica oleracea&nbsp; </em>i<em>&nbsp; Brassica rapa</em>&nbsp; poreklom iz organskog i konvencionalnog sistema proizvodnje. Ispitivanja hemijskog sastava su obuhvatila određivanje volatilnih komponenata primenom headspace GC-MS tehnike, kvalitativnu analizu sokova ispitivanih vrsta pomoću LC-DAD-MS/MS tehnike, kvantifikaciju odabranih fenolnih jedinjenja&nbsp;pomoću LC-MS/MS tehnike i spektrofotometrijsko određivanje sadržaja ukupnih glukozinolata, fenola i flavonoida i sadržaja ukupnih monomernih antocijana. Biolo&scaron;ka aktivnost povrća je ispitana određivanjem antioksidantnog potencijal, antimikrobne i antitumorske aktivnosti sokova. Dobijeni rezultati pokazuju da su glavne volatilne komponente ispitivanih vrsta degradacioni proizvodi masnih kiselina i organosumpornih jedinjenja od kojih neke od identifikovanih komponenata predstavljaju ključne mirisne komponente. Na osnovu njihove distribucije nije bilo moguće utvrditi uticaj sistema proizvodnje na miris i ukus ispitivanih vrsta. Pomoću LC-DAD-MS/MS i LC-MS/MS tehnike je utvrđeno prisustvo primarnih metabolita, slobodnih hidroksibenzoevih i hidroksicimetnih kiselina, derivate hinske kiseline&nbsp;sa hidroksicimetnim kiselinama, glikozida flavonola, biflavonoida, i kumarina u sokovima spitivanih vrsta. Pored toga u svim sokovima subili prisutni kondenzacioni proizvodi degradacionih proizvoda indolnih glukozinolata sa askorbinskom kiselinom-&nbsp; askorbigen i metoksiaskorbigen. U sokovima crvenog kupusa su bili prisutni antocijani. Najveći sadržaj ukupnih glukozinolata je određen u brokoliju, dok je crveni kupus sadržao najveći sadržaj ukupnih fenola ukupnih flavonoida kao i hinske kiseline. Crveni kupus je bio varijetet koji je ispoljio najvi&scaron;i antioksidantni kapacitet u svim primenjenim testovima, brokoli i karfiol su ispoljili najjaču antimikrobnu aktivnost, dok je kelj bio varijetet sa najjačim antitumorskim delovanjem. Na osnovu analize hemijskog sastava i biolo&scaron;ke aktivnosti vrsta <em>B</em>. <span id="cke_bm_177S" style="display: none;">&nbsp;</span><em>oleracea</em><span id="cke_bm_177E" style="display: none;">&nbsp;</span> i&nbsp;<em> B. rapa</em>&nbsp; može se <span id="cke_bm_178E" style="display: none;">&nbsp;</span>zaključiti da u zavisnosti od primenjenog tipa&nbsp;proizvodnje mogu biti pobolj&scaron;ane neke od karakteristika ispitivanih vrsta i da je uticaj sistema proizvodnje značajan za pojedine varijetete, a nikako za celu vrstu. U konačnom obliku može se zaključiti da bi potencijalna kombinacija sokova crvenog kupusa, brokolija, karfiola i kelja mogla ostvariti značajno hemopreventivno i hemoterapeutsko delovanje, ispoljavajući istovremeno antioksidantnu, antimikrobnu i antitumorsku aktivnost.</p> / <p>Chemical composition and biological activity of<em>&nbsp; Brassica oleracea</em>&nbsp; and&nbsp; <em>B. rapa&nbsp;</em> species were investigated in the present work. Investigation of chemical composition&nbsp; encompassed determination of volatile compounds by headspace GC-MS technique, qualitative analysis of juices of investigated vegetables by LC-DAD-MS/MS technique, quantification of selected phenolic compounds by LC-MS/MS technique and spectrophotometric determination of total content of glucosinolates, phenolic compounds and flavonoids and total content of monomeric anthocyanes. Biological activity of vegetables was investigated by determination of antioxidant capacity, antimicrobial and antitumorigenic effect of juices. Obtained results showed that main volatile compounds were degradation products of fatty acids and organo-sulphur compounds among which some of the identified components were the key aroma compounds. Based on their distribution it was not possible to evaluate influence of cropping system on aroma and taste of investigated species. By LC-DAD-MS/MS and LC-MS/MS techniques, it was determined the presence of primary metabolites, freehydroxybenzoic and hydroxycinnamic acids, derivatives of quinic acid and hidroxicinnamic acids, flavonol glucosides, biflavnoids&nbsp; and a cumarin in juices of the investigated species. Furthermore, condensation products of degradation products of indole glucosinolates with ascorbic acid-&nbsp; ascorbigen and metoxiascorbigen were present in all juices. Anthocyanes were present in juices of red cabbage. The highest total content of glucosinolates was determined in broccoli, while red cabbage had the highest total phenolic content, total flavonoid content, as well as content of qunic acid. Red cabbage was a variety that showed the highest antioxidant capacity measured by different assays. Broccoli and cauliflower expressed the highest antimicrobial potential, while Savoy cabbage was a variety with the highest antitumor effect. Investigation of chemical composition and biological activity showed that applied cropping conditions may be useful for improving some of the characteristics of investigated species and significance of cropping system only&nbsp; for particular varieties, but not for the whole species in general. As the conclusion, potential&nbsp; combination of juices of red cabbage, broccoli, cauliflower and Savoy cabbage could be potent chemopreventive and chemotherapeutic agent, expressing simultaneously antioxidant, antimicrobial and antitumor activity.</p>

<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>

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Genus Senecio (Asteraceae) constitutes a group of cosmopolitan plants formed by more than 2000 species. Although these plants have toxicity recognized due to the presence of pyrrolizidine alkaloids, several of them are used in folk medicine. Their medicinal use can be attributed to other secondary metabolites such as flavonoids and terpenoids, being that terpenoids are compounds with recognized antimicrobial activity. In this work, the evaluation of the antimicrobial activity of the CH2Cl2 and EtOH extracts of S. heterotrichius DC., the isolation and characterization of one constituent from CH2Cl2 extract, the reaction of photooxidation of this compound in order to obtain a peroxide as well as the chemical analysis and evaluation of the antimicrobial activity of the essential oils from Senecio heterotrichius DC., S. ceratophylloides Griseb., S. oxyphyllus DC. and S. leptolobus DC. were studied. Extracts of the fresh aerial parts of S. heterotrichius were obtained by sequential maceration at room temperature in CH2Cl2 and EtOH, followed by evaporation under reduced pressure. The CH2Cl2 extract was firstly fractioned by CC over silica gel for the isolation of the sesquiterpene germacrene D and then identified by GC-MS, 1H and 13C NMR. This compound, considered a precursor of several other sesquiterpene derivatives, was submitted to photooxidation reaction using Bengal rose as sensitizer agent. The main formed peroxide was isolated from the reactional mixture by CC over silica gel, analyzed by GC-MS,1H and 13C NMR, and identified as being germacrene-D-1-hydroperoxide. The evaluation of the antimicrobial activity of the extracts of S. heterotrichius and of germacrene-D-1-hydroperoxide was accomplished through the broth microdilution method based on M27- A2/NCCLS documents for yeasts and M7-A4/NCCLS for aerobic bacteria. The extraction of the essential oils of the flowery aerial parts of Senecio heterotrichius, S. ceratophylloides, S. oxyphyllus and S. leptolobus was accomplished through hydrodestilation. The obtained essential oils were analyzed by GC-MS and their constituents were identified by the comparison of their Kovat s Indexes and their fragmentation patterns with literature data. These oils also had their antimicrobial activity evaluated following the previously mentioned methodology. The CH2Cl2 extract evidenced inhibitory activity against Candida krusei (MIC of 0.25 mg/mL) and Staphylococcus aureus (MIC of 2.5 mg/mL). The EtOH extract was inactive against the tested microorganisms. The germacrene-D-1-hydroperoxide presented inhibitory activity against S. cerevisiae (MIC of 2.5 mg/mL), and inhibited partially the growth of the algae Prototheca zoopfi at 5 and 2.5 mg/mL (inhibition of 80 and 50%, respectively). The essential oils of S. ceratophylloides and S. leptolobus exhibited inhibitory and bactericidal activity against Staphylococcus aureus and Pseudomonas aeruginosa (MIC and MBC between 6.6 and 3.3 mg/mL). The oil of S. oxyphyllus just inhibited partially the growth of these microorganisms (MIC 50% = 6.6 and 1.6 mg/mL, respectively). The oil of S. heterotrichius inhibited partially the growth of S. aureus with a MIC 50% = 3.3 mg/mL. These oils are constituted almost exclusively by mono and sesquiterpenoids, being germacrene D one of the main constituents present in all essential oils. / O gênero Senecio (Asteraceae) constitui um grupo de plantas cosmopolitas formado por mais de 2000 espécies. Embora estas tenham toxicidade reconhecida devido à presença de alcalóides pirrolizidínicos, várias delas são empregadas na medicina popular. Seu uso medicinal pode ser atribuído aos demais metabólitos secundários, entre eles flavonóides e terpenóides, sendo estes últimos, compostos com reconhecida atividade antimicrobiana. Neste estudo é relatada a avaliação da atividade antimicrobiana dos extratos CH2Cl2 e EtOH de S. heterotrichius DC., o isolamento e caracterização de um constituinte do extrato CH2Cl2, a reação de fotooxidação deste constituinte obtendo-se um peróxido, bem como a análise química e avaliação da atividade antimicrobiana dos óleos essenciais de Senecio heterotrichius DC., S. ceratophylloides Griseb., S. oxyphyllus DC. e S. leptolobus DC. Os extratos das partes aéreas frescas de S. heterotrichius foram obtidos por maceração seqüencial à temperatura ambiente em CH2Cl2 e EtOH, seguido de evaporação sob pressão reduzida. O extrato CH2Cl2 foi fracionado por CC sobre gel de sílica até o isolamento do germacreno D, identificado por CG-EM, RMN 13C e 1H. Este composto, considerado um precursor de vários outros derivados de esqueleto sesquiterpênico, foi submetido a uma reação de foto-oxidação utilizando o corante Rosa de Bengala como agente sensibilizante; o principal peróxido formado foi isolado da mistura reacional por CC sobre gel de sílica, analisado por CG-EM, RMN 13C e 1H, e identificado como sendo o germacreno-D-1-hidroperóxido. A avaliação de atividade antimicrobiana dos extratos de S. heterotrichius e do germacreno-D-1-hidroperóxido foi realizada através do método de microdiluição em caldo, baseado nos documentos M27-A2/NCCLS para fungos leveduriformes e M7-A4/NCCLS para bactérias aeróbias. A extração dos óleos essenciais das partes aéreas floridas de Senecio heterotrichius, S. ceratophylloides, S. oxyphyllus e S. leptolobus foi realizada pela técnica de hidrodestilação. Os óleos essenciais obtidos foram analisados por CG-EM e seus constituintes foram identificados através da comparação de seus Índices de Kovat s e dos modelos de fragmentação com dados da literatura. Estes óleos também tiveram sua atividade antimicrobiana avaliada conforme metodologia citada anteriormente. O extrato CH2Cl2 evidenciou atividade inibitória frente Candida krusei (CIM de 0,25 mg/mL) e Staphylococcus aureus (CIM de 2,5 mg/mL). Já o extrato EtOH foi inativo frente os microrganismos testados. O germacreno-D-1-hidroperóxido apresentou atividade inibitória frente S. cerevisiae (CIM de 2,5 mg/mL) e inibiu parcialmente o crescimento da alga Prototheca zoopfi nas concentrações de 5 e 2,5 mg/mL (80 e 50 % de inibição, respectivamente). Os óleos essenciais de S. ceratophylloides e S. leptolobus apresentaram atividade inibitória e bactericida frente Staphylococcus aureus e Pseudomonas aeruginosa (CIM e CBM entre 6,6 e 3,3 mg/mL); o óleo de S. oxyphyllus apenas inibiu parcialmente o crescimento destes microrganismos (CIM 50% = 6,6 e 1,6 mg/mL, respectivamente); já o óleo de S. heterotrichius inibiu parcialmente o crescimento de S. aureus com uma CIM 50% = 3,3 mg/mL. Estes óleos são constituídos quase que exclusivamente de mono e sesquiterpenóides, sendo o germacreno D um dos constituintes majoritários presentes em todos os óleos essenciais.

No presente trabalho, destinado a tese de doutorado, desenvolvemos um estudo da relação entre a estrutura química e atividade antimicrobiana de duas séries, a saber: Série I - quatorze cloretos N,N[(dimetilamino)etil]-benzoatos para-substituídos, onde R4 = H, CH3, C2H5, nC4H9, tC4H9, OCH3, nOC4H9, COCH3, Cl, CN, NO2, OC6H13, SO2Me, CF3 (compostos I.1 a I.14) e Série II - quatorze cloretos meta-substituídos onde R3 = H, -CH3, -OCH3, -OC2H5, -OC4H9, -F, -Cl, -Br, -I, -NO2, -CN, -CF3, -N(CH3)2, SO2CH3 (compostos II.1. a II.14). Os compostos haviam sido sintetizados anteriormente no grupo (Amaral et al, 1997; Sousa, 1997). Para as séries I e II, os valores do coeficiente de partição em n-octanol foram obtidos por cálculo (utilizando os programas CLOGP e ACD/LogP) bem como determinados experimentalmente pelo método CLAE e considerados parâmetros lipofílicos nos modelos de QSAR. Adicionalmente, para as séries I e II, os valores da posição da banda de absorção da carbonila na região do infravermelho bem como do deslocamento químico da carbonila no RMN 13C foram determinados sendo o primeiro considerado como parâmetro eletrônico nos modelos QSAR. E, ainda, para as séries I e II os valores de MR3 e de MR4 , relacionados à refratividade molar, foram obtidos da literatura e considerados nas análises QSAR . Para as séries I e II os valores de IC50 (concentração inibição 50% do crescimento) foram determinados contra, respectivamente, o fungo Saccharomyces cerevisiae (BY4741) e contra a bactéria Escherichia coli (DH5&#945;) e considerados como parâmetros biológico nas análises QSAR. Para as séries I e II e para as séries I e II reunidas, foram gerados modelos QSAR e estes foram analisados visando avaliar a natureza e a contribuição de cada parâmetro estrutural para cada uma das atividades avaliadas. A aplicação do modelo linear resultou em modelos iestatisticamente significativos para as atividades contra o Saccharomyces cerevisiae (BY4741) e contra a Escherichia coli (DH5&#945;) para a série I e para as séries I e II reunidas. A série II não gerou modelos lineares estatisticamente significativos. A aplicação dos modelos bilinear e parabólico foram verificadas não resultando em modelos estatisticamente significativos. Para a série I e para as séries I e II reunidas estudadas neste trabalho, as diferenças observadas nos modelos QSAR gerados para a inibição do crescimento de, respectivamente, S. cerevisiae (BY4741) e de E. coli (DH5&#945;) podem ser, possivelmente, explicadas considerando-se as diferenças nas membranas externas de cada microorganismo. / In this PhD thesis work, we developed a quantitative structure relationship study for the antimicrobial activity of two series, namely: Series I fourteen para-substituted N,N- [(dimethylamino)ethyl)] benzoates chlorides, where R4 = -H, -CH3, -C2H5, -nC4H9, -tC4H9, -OCH3, - nOC4H9, -COCH3, -Cl, -CN, -NO2, -OC6H13, -SO2Me, -CF3 (compounds I.1 - I.14) and Series II - fourteen meta-substituted chlorides, where R3 = -H, -CH3, -OCH3, -OC2H5, -OC4H9, -F, -Cl, -Br, -I, -NO2, -CN, -CF3, -N(CH3)2, -SO2CH3 (compounds II.1. - II.14). These compounds had been previously synthesized in the group (Amaral et al, 1997; Sousa, 1997). For series I and II, the partition coefficient values in n-octanol were obtained by calculation (using the CLOGP and the ACD/LogP programs) and were obtained experimentally by HPLC method. These values were taken as lipophilic parameters in the QSAR analysis. In addition, for the series I and II, the carbonyl infrared absorption band values and 13C-NMR chemical shifts of carbonyl group were determined, being the former considered as electronic parameters in the QSAR analysis. Further, for the series I and II, the MR3 and MR4 values were taken from the literature and considered as molar refractivity related parameter in the QSAR analyses. For the series I and II, IC50 values (concentration inhibiting 50% growth) were determined against, respectively, Saccharomyces cerevisiae (BY4741) yeast and Escherichia coli (DH5&#945;) bacteria and taken as biological parameters in the QSAR analysis. For the series I and II and for series I e II altogether, QSAR models were generated and analysed in order to determine the nature and the contribution of each structural parameter to the determined antimicrobial activities. The application of linear model resulted in statistically significant QSAR models for the activities evaluated against Saccharomyces cerevisiae (BY4741) and against Escherichia coli (DH5&#945;) for series I and for series I and II taken altogether. No statistically significant linear models were obtained for series II. The application of bilinear and parabolic models was verified, resulting in no statistically significant models. For series I and series I e II altogether, the differences observed in the generated QSAR models for the inhibition, respectively, of Saccharomyces cerevisiae (BY4741) and of E. coli (DH5&#945;) growth, can be, probably, explained considering the differences in the external membranes of each microorganism.

Made available in DSpace on 2014-07-29T15:10:33Z (GMT). No. of bitstreams: 1 Tese M nodosa Final I.pdf: 3081089 bytes, checksum: 1751ddbcf974cb6b37ba7e8b90043507 (MD5) Previous issue date: 2008-12-18 / Memora nodosa (Silva Manso) Miers is a native savannah plant popularly used for wound cicatrization (leaves and stems) and for treating abdominal pains and scabies (roots). The purpose of this work is to study the morphoanatomy and determine the composition and variability of the constituents of the essential oil obtained from the M. nodosa leaves, as well as its antimicrobial, leishmanicidal, cicatrizing activity and toxicity. Morphologically, M. nodosa is a bush with clumps of straight or curved stems. It has a deep, pivoting root and presents a chandelier shaped structure in the upper part from where the stems sprout. It presents compound, bipinnate, imparipinnate, oppositely crossed leaves. The inflorescence is racemose with golden-yellow flowers. It is characterized by septifrague capsule type fruit with replum and winged seeds. Anatomically the leaves are hypostomatic with predominantly paracytic stomates. The epidermis is uniseriate with a thick cuticle and glandular trichomes throughout the structure of the young leaves. The essential oil from the M. nodosa leaves collected from seven different locations in the Brazilian savannah was analyzed by GC(FID) and GC/MS. Five substances were identified, benzaldehyde and 1- octene-3-ol were the main constituents. Two different classes regarding sampling location and constituents were identified by means of the principal component and grouping analyses. The antimicrobial activity of the ethanolic extracts from leaves, stems, roots and of essential oil from leaves, and the fractions obtained from the roots, were assessed against Grampositive and Gram-negative bacteria, as well as the Candida albicans fungus, using well diffusion test and the agar dilution method. The ethanolic extract from the roots presented small antimicrobial activity while the hexane and dichloromethane fractions obtained from this extract presented MIC 0.39 mg/mL and 0.78 mg/mL against a number of Gram-positive bacteria and the Candida albicans fungus. The essential oil presented MIC 0.78 mg/mL to 1.56 mg/mL. The antimicrobial activity of the essential oil was the result of synergic action between the benzaldehyde, 1-octene-3-ol, 1-octanol, linalool and the mandelonitrile, constituents of the essential oil. Promastigote forms of Leishmania chagasi were used for the leishmanicidal assessment and it was verified that the ethanol extract from the leaves presented an IC50 of 93.2 &#956;g/mL and from the roots presented an IC50 of 95.87 &#956;g/mL. The hexane fraction from the roots presented an IC50 of 13.51&#956;g/mL. A mixture of &#946;-sitosterol and stigmasterol was isolated and identified in the hexane fraction obtained from the ethanol extract from the roots. This mixture was tested against L. chagasi resulting in an IC50 of 69.79 &#956;g/mL. The cicatrizing activity of the ethanol extract from the leaves and roots was assessed on skin wounds in rats, and it be noted reepithelization occurred faster only in the group treated with a 2% aqueous solution of the ethanol extract from the roots. This action may be related to the alantoine isolated in the ethanol extract from the roots. The toxicity assessment of the ethanol extracts from roots, leaves and hexane fraction obtained from M . nodosa roots, tested on rats, mice and Artemia salina proved that this plant is not potentially toxic. In conclusion, according to the tests performed, the best results were obtained with ethanolic extract and it presented antimicrobial, wound cicatrization and leishmanicidal activities. Hexane and dichloromethane fractions obtained from roots extract presented antimicrobial activity against Gram-positive bacteria and the Candida albicans fungus and hexane fraction strong leishmanicidal activity. / A Memora nodosa (Silva Manso) Miers é uma planta nativa do Cerrado e utilizada popularmente na cicatrização de feridas (folhas e caules) e no tratamento de dores abdominais e sarnas (raízes). Este trabalho objetivou realizar o estudo morfoanatômico, determinar a composição química e a variabilidade dos constituintes do óleo essencial obtido das folhas e avaliar a atividade antimicrobiana, leishmanicida e cicatrizante e a toxicidade dos extratos e frações da M. nodosa. Morfologicamente, observou-se que a M. nodosa é um arbusto com caules retos ou escandentes reunidos em touceiras. A raiz é pivotante, profunda e apresenta, na parte superior, uma estrutura em candelabro de onde se originam os caules. As folhas são opostas cruzadas, recompostas, bipinadas, imparipinadas, com foliólulos lanceolados. A inflorescência é racemosa com flores de cor amarelo-ouro. O fruto é do tipo cápsula septífraga com abertura longitudinal e sementes aladas. Anatomicamente, as folhas são hipoestomáticas, com estômatos predominantemente paracíticos. A epiderme é unisseriada, com cutícula espessa em todas as estruturas das folhas. Tricomas glandulares e tectores são observados nas folhas e caules jovens. O óleo essencial das folhas da M. nodosa coletadas em sete localidades diferentes do Cerrado brasileiro foi analisado por CG (DIC) e CG/EM. Cinco substâncias foram identificadas, sendo o benzaldeído e o 1-octen-3-ol os principais constituintes. A análise dos componentes principais e dos agrupamentos possibilitou a distinção de duas classes diferenciadas em relação ao local da amostragem e aos constituintes. As atividades antimicrobianas dos extratos etanólicos das folhas, dos caules e das raízes, do óleo essencial das folhas e das frações obtidas das raízes foram avaliadas usando teste de difusão em poço e o método de diluição em ágar. O extrato etanólico das raízes apresentou atividade antimicrobiana fraca, enquanto as frações hexano e diclorometano, obtidas desse extrato, mostraram atividade de forte a moderada contra várias bactérias Gram-positivas e contra o fungo Candida albicans. O óleo essencial obtido das folhas apresentou CIM de 0,78 mg/mL a 1,56 mg/mL contra bactérias Gram-positivas e Candida albicans. Constatou-se que a atividade antimicrobiana do óleo essencial se deve a ação sinérgica entre o benzaldeído, o 1- octen-3-ol, o 1-octanol, o linalol e a mandelonitrila, constituintes do óleo essencial. A atividade cicatrizante dos extratos etanólicos das folhas e raízes foi avaliada em feridas cutâneas de ratos e observou-se uma aceleração na reepitelização apenas no grupo tratado com solução aquosa a 2% do extrato etanólico das raízes. Essa ação pode estar relacionada com a alantoína, isolada do extrato etanólico das raízes. Na avaliação da atividade leihsmanicida, empregaram-se culturas com formas promastigotas de Leishmania chagasi e verificou-se que o extrato etanólico das folhas apresentou IC50 de 93,2 &#956;g/mL e as raízes IC50 de 95,87 &#956;g/mL. A fração hexano obtida das raízes apresentou IC50 de 13,51 &#956;g/mL. Da fração hexano obtida do extrato etanólico das raízes foi isolado e identificado uma mistura de &#946;-sitosterol e estigmasterol. Essa mistura foi testada contra L. chagasi, obtendo-se IC50 de 69,79 &#956;g/mL. A avaliação da toxicidade aguda dos extratos etanólicos das folhas, das raízes e da fração hexano obtidas das raízes empregando como modelos experimentais ratos e camundongos e o teste de citotoxicidade utilizando Artemia salina mostraram que essa planta não é potencialmente tóxica. Conclui-se que os melhores resultados foram obtido com extrato etanólico das raízes, que apresentou atividade antimicrobiana, cicatrizante e leishmanicida.As frações hexano e diclorometano, obtidas das raízes, apresentaram bom potencial antimicrobiano contra bactérias Gram-positivas e o fungo C. albicans e a fração hexano forte atividade leishmnicida.

<p>Испитивано је хроматографско понашање (хроматографска липофилност) 29 стероидних једињења (триазола и тетразола, толуенсулфонилхидразида, диона, нитрила и динитрила) од потенцијалног биомедицинског значаја, испитивано је помоћу течне хроматографије високих перформанси на обрнутим фазама, применом две стационарне и две мобилне фазе. Липофилност, изражена преко ретенционог параметра logk, моделована је QSRR приступом. Формирани линеарни и нелинеарни модели омогућили су испитивање односа између ретенционих параметара и in silico молекулских дескриптора, који су израчунати на основу структуре испитиваних једињења. Добра предиктивна моћ формираних модела, добијених за калибрациони сет, потврђена је и применом екстерног тест сета и валидационог сета. Предиктивна моћ формираних модела потврђује могућност њиховог коришћења за предвиђање липофилности нових, структурно сличних, једињења. Примењене су и класификационе хемометријске методе (анализа главних компоненти и хијерархијска кластер анализа) како би се уочиле сличности и разлика између једињења. Поред тога, представљена је in vitro анализа антимикробног потенцијала испитиваних стероидних једињења према Staphylococcus aureus, Escherichia coli и Candida albicans. Два једињења, са епоксидном групом у положају 4,5, испољила су бактериостатски ефекат према S. aureus. Такође, приказана је докинг анализа одабраних испитиваних једињења са антипролиферативном активношћу према ћелијама андроген-рецептор негативног канцера простате (AR-нег. PC-3). На основу визуелизације оптималних положаја и анализе постојећих интеракција, идентификовано је једињење са највећим потенцијалом као инхибитор хуманог цитохрома P450 CYP17A1.</p> / <p>Ispitivano je hromatografsko ponašanje (hromatografska lipofilnost) 29 steroidnih jedinjenja (triazola i tetrazola, toluensulfonilhidrazida, diona, nitrila i dinitrila) od potencijalnog biomedicinskog značaja, ispitivano je pomoću tečne hromatografije visokih performansi na obrnutim fazama, primenom dve stacionarne i dve mobilne faze. Lipofilnost, izražena preko retencionog parametra logk, modelovana je QSRR pristupom. Formirani linearni i nelinearni modeli omogućili su ispitivanje odnosa između retencionih parametara i in silico molekulskih deskriptora, koji su izračunati na osnovu strukture ispitivanih jedinjenja. Dobra prediktivna moć formiranih modela, dobijenih za kalibracioni set, potvrđena je i primenom eksternog test seta i validacionog seta. Prediktivna moć formiranih modela potvrđuje mogućnost njihovog korišćenja za predviđanje lipofilnosti novih, strukturno sličnih, jedinjenja. Primenjene su i klasifikacione hemometrijske metode (analiza glavnih komponenti i hijerarhijska klaster analiza) kako bi se uočile sličnosti i razlika između jedinjenja. Pored toga, predstavljena je in vitro analiza antimikrobnog potencijala ispitivanih steroidnih jedinjenja prema Staphylococcus aureus, Escherichia coli i Candida albicans. Dva jedinjenja, sa epoksidnom grupom u položaju 4,5, ispoljila su bakteriostatski efekat prema S. aureus. Takođe, prikazana je doking analiza odabranih ispitivanih jedinjenja sa antiproliferativnom aktivnošću prema ćelijama androgen-receptor negativnog kancera prostate (AR-neg. PC-3). Na osnovu vizuelizacije optimalnih položaja i analize postojećih interakcija, identifikovano je jedinjenje sa najvećim potencijalom kao inhibitor humanog citohroma P450 CYP17A1.</p> / <p>Chromatographic behavior (chromatographic lipophilicity) of 29 steroid compounds (triazole and tetrazole, toluenesulfonylhydrazide, dione, dinitrile and nitrile) with potential biomedical importance was investigated by reversed-phases high-performance liquid chromatography using two stationary and two mobile phases. The lipophilicity expressed through the retention parameter logk was modeled using QSRR approach. Formed linear and non-linear models enabled the study of the relationship between the retention parameters and in silico molecular descriptors calculated from the structure of the investigated compounds. Good predictive power of the established models obtained for the calibration set was confirmed by the application of an external test set and validation set. The predictive power of the established model confirms the possibility of their use for lipophilicity prediction of new, structurally similar compounds. The classification chemometric methods (principal components analysis and hierarchical cluster analysis) were applied in order to recognize the similarities and differences between the compounds. Тhis dissertation presents the in vitro analysis of the antimicrobial potentials of the investigated steroid compounds against Staphylococcus aureus, Escherichia coli and Candida albicans. Two compounds, with epoxy group in the position 4,5, exhibited bacteriostatic effect against S. aureus. The docking analysis of selected test compounds with antiproliferative activity toward cells of androgen receptor-negative prostate cancer (AR-neg. PC-3) is showed. Based on the optimal position visualization and analysis of existing interactions a compound with the most promising potential as human cytochrome P450 CYP17A1 inhibitor is idetified.</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>U okviru ove disertacije ispitan je hemijski, mineralni i<br />polifenolni sastav, kao i biolo&scaron;ki potencijal četiri sorte<br />kupina sa dva različita lokaliteta sjeverozapadnog dijela<br />Bosne i Hercegovine, sa lokaliteta Verići (divlja i<br />kultivisana sorta Čačanska bestrna) i sa lokaliteta<br />Javorani (divlja sorta i kultivisana sorta Chester<br />Thornles). Određen je sadržaj suve materije, pepela,<br />sirove celuloze, ukupnih &scaron;ećera, ukupna kiselost i sadržaj<br />askorbinske kiseline. Sadržaj mineralnih materija<br />detektovan je metodom optičke emisione spektrometrije<br />(ICP-OES). Za izdvajanje ekstrakata je kori&scaron;tena klasična<br />ekstrakciona tehnika prema Soxhlet-u uz upotrebu 80 %<br />etanola (v/v). Spektrofotometrijskim metodama određen<br />je sadržaj ukupnih polifenola, flavonida, flavonola,<br />ukupnih i monomernih antocijana. Kvalitativnom i<br />kvantitativnom HPLC metodom je utvrđen sadržaj<br />pojedinačnih polifenolih jedinjenja. Biolo&scaron;ki potencijal<br />uzoraka je utvrđen u sistemima in vitro, gdje je određena:<br />antioksidativna aktivnost, antihiperglikemijska aktivnost,<br />antiproliferativni efekat i antimikrobna aktivnost.<br />Antioksidativna aktivnost uzoraka je ispitana sa četiri<br />metode: DPPH test, ABTS test, sposobnost neutralizacije<br />hidroksil radikala i metodom inhibicije Briggs Rauscherovih oscilatornih reakcija. Antihiperglikemijska aktivnost<br />ispitivanih uzoraka kupine je dokazan na osnovu<br />sposobnosti inhibicije enzima &alpha;-glukozidaze.<br />Antiproliferativni efekat ispitivanih uzoraka je određen<br />prema inhibiciji rasta četiri humane ćelijske linije: HeLa<br />(epitelnog karcinoma cerviksa), HT-29 (adenokarcinoma<br />debelog crijeva), MRC-5 (zdravim ćelijama fibroblasti<br />pluća) i prema ćelijskoj liniji MCF7 (adenokarcinoma<br />dojke). Antimikrobna aktivnost je utvrđena prema gram<br />pozitivnom soju (G⁺) bakterija S. Aureus i gram<br />negativnom (G^-) soju E. coli, rastu micelija Aspergillusa<br />niger i Candida albicans. Regresionom analizom prema<br />Pirson-u su određeni odnosi između sadržaja polifenolnih<br />jedinjenja i biolo&scaron;kog potencijala, sa statističkim<br />značajem (p &le; 0,01).</p> / <p>The chemical, mineral, polyphenolics composition and<br />biological potentials of four blackberries varieties from two<br />different locations in the northwestern part of Bosnia and<br />Herzegovina (Verići - wild and cultivated variety Čačanska<br />bestrna and Javorani - wild and cultivated variety Chester<br />Thornless) were determined. The contents of dry matter, ash,<br />crude cellulose, total sugars, total acidity and ascorbic acid were<br />obtained. The contents of mineral matter were detected by<br />optical emission spectrometry (ICP-OES) method. A classic<br />Soxhlet extraction technique with 80% ethanol (v/v) was<br />applied to obtain extracts. The spectrophotometric methods<br />were used to determine the content of total polyphenolics,<br />flavonoids, flavonols, and total and monomer anthocyanins. The<br />content of individual polyphenolic compounds was determined<br />by HPLC method. The biological potentials (antioxidant<br />activity, antihyperglycemic activity, antiproliferative effect and<br />antimicrobial activity) of the samples were determined in vitro<br />systems. The antioxidant activity was tested with four methods:<br />DPPH test, ABTS test, ability to neutralize OH radicals and the<br />method of inhibiting Briggs Rauscher oscillatory reactions. The<br />antihyperglycaemic activity of the tested blackberry samples<br />was based on the ability to inhibit &alpha;-glucosidase enzyme. The<br />antiproliferative effect of the tested samples was determined by<br />inhibiting the growth of four human cell lines: epithelial<br />carcinoma of the cervix (HeLa), colon adenocarcinoma (HT-<br />29), healthy lung fibroblast cells (MRC-5) and the cell line of<br />breast adenocarcinoma (MCF7). The extracts showed the<br />highest inhibitory effect on the cell line of breast<br />adenocarcinoma (MCF-7). The antimicrobial activity was<br />determined according to gram-positive bacteria (G ⁺) of<br />Staphylococcus aureus, Gram-negative bacteria (G ^-)<br />Escherichia coli, growth of mycelium Aspergillus niger and<br />fungi Candida albicans. The Pearson correlations were<br />statistically determined the relationship between the content of<br />polyphenolic compounds and biological potential, with<br />statistical significance (p &le; 0.01).</p>

The presented doctoral thesis is focused on preparation, characterization and application of organic micro- and nanoparticles as transport systems for active components and some their complex natural sources. Active component were packed into liposomes and polysaccharide particles. As active components were used caffeine, some drugs – clotrimazole and ibuprofen, further antioxidants and vitamins. Antimicrobial herbs and spices extract, antimicrobial peptides lysozyme, nisin and other antimicrobial ingredients were encapsulated too. Encapsulation of selected hydrolytic enzymes was tested, too. Particles were also used for encapsulation of probiotic strains Bifidobacterium breve and Lactobacillus acidophilus and prebiotic components. These prebiotics were co-encapsulated into capsules with probiotic cells. Natural extracts were encapsulated e.g. extracts of guarana, ginseng, goji, green barley, propolis, black, green and white tea, coffee, fruit and vegetable extracts. The efficiency of encapsulation was determined by HPLC/PDA and by spectrophotometry. Long-term stability of particles and amount of released component in model/real foods, in model cosmetic conditions and in a model physiological environment were monitored too. Size of prepared liposomes and polysaccharide particles was determined by dynamic light scattering and by light microscopy and electron microscopy, respectively. Stability of the particles was measured using a zeta potential. Also, analytical centrifugation was used to measurement of sedimentation velocity and stability of the prepared particles. The antimicrobial activity were tested using two Gram-positive (Bacillus subtilis, Micrococcus luteus), two Gram-negative (Escherichia coli, Serratia marcescens) bacteria and one fungal strains (Candida glabrata). For determining the antimicrobial properties of active component and prepared particles two the most widely used methods were used - agar diffusion method and broth dilution method. The viability of probiotic strains were performed using flow cytometry and fluorescence microscopy. Encapsulation of active component was successful in all types of particles. Liposome showed a very good long-term stability mainly in water conditions with neutral pH and polysaccharide particles were stable in acidic conditions. Prepared particles showed a very good stability in model stomach environment, while in model intestines environments particles were disintegrated and active component were released. Prepared particles with encapsulated caffeine as well as other tested antioxidants and vitamins could be used to modern types of energy drinks, food supplements and also for some cosmetics applications. Encapsulated antimicrobial components could be used for food application as well as for cosmetics and pharmaceutical application like antimicrobial wound formulation. Encapsulated enzymes can be used for controlled release of proteases in wound healing, as delivery systems in digestive tract and as a part of pharmaceutical preparative and food supplements for enzyme therapy. The study revealed that encapsulation of probiotics and also co-encapsulation of probiotics with prebiotics exhibited longer stability of particles and survival bacterial cells. So, prepared particles are suitable for use to food product with beneficial effects on the human body.

<p>Hemijska karakterizacija ekstrakata&nbsp; <em>T. pratense&nbsp;</em>L.&nbsp; određena je LC-MS-MS, a etarskih ulja GC-MS analizom, pri čemu je&nbsp; identifikovano vi&scaron;e&nbsp;bioaktivnih jedinjenja.&nbsp; Spektrofotometrijskim&nbsp;metodama ispitan je antioksidantni potencijal&nbsp;ekstrakata i etarskih ulja. Hepatoprotektivni&nbsp;efekat određen je&nbsp; na&nbsp; homogenatu i hemolizatu&nbsp;jetre laboratorijskih mi&scaron;eva.&nbsp; Budući da&nbsp; ova&nbsp;biljka ima visok sadržaj izoflavona, određena je&nbsp;<br />estrogena, antiestrogena aktivnost, kao i&nbsp;citotoksičnost ekstrakata. Antimikrobna&nbsp;<br />aktivnosta ekstrakata i etarskih ulja ispitana je&nbsp;na&nbsp; &scaron;est bakterijskih sojeva.&nbsp; Ekstrakti i etarska&nbsp;ulja su bili umereno aktivni u pogledu biolo&scaron;kih&nbsp;aktivnosti, osim u slučaju antimikrobne&nbsp;aktivnosti koju nije pokazao nijedan ispitivani&nbsp;uzorak.</p> / <p>The chemical characterization of&nbsp; <em>T.&nbsp; pratense</em> extracts was determined by LC-MS-MS, and the essential oils of the GC-MS analysis, wherein the&nbsp; various&nbsp; bioactive compounds were identified.&nbsp; Antioxidant potential of extracts and essential oils was tested&nbsp; by spectrophotometric methods.&nbsp; Hepatoprotective&nbsp; effect&nbsp; was determined&nbsp; in&nbsp; haemolysate and liver homogenate of&nbsp; laboratory&nbsp; mice.&nbsp; Estrogen, antiestrogen activity, as well&nbsp; as the cytotoxicity of the extracts were&nbsp; determined&nbsp; due to the high&nbsp;content of isoflavones.&nbsp; Antimicrobial activity of extracts and essential oils was&nbsp; investigatied&nbsp; on six bacterial strains. Essential oils and extracts were moderately active in terms of biological activity, except&nbsp; in the case of&nbsp;&nbsp; antimicrobial activity&nbsp; where none of the extracts were active against tested bacterial strains.</p>