En période néonatale, la muqueuse intestinale est immature et particulièrement perméable, notamment aux facteurs environnementaux comme les toxiques, les infections mais aussi les événements stressants. De la bonne maturation de la barrière intestinale va dépendre la mise en place de l'homéostasie intestinale et donc la tolérance vis-à-vis des antigènes luminaux. La survenu d’évènements stressants durant la petite enfance est associée au déclenchement et/ou l’entretien de pathologies gastro-intestinales fonctionnelles comme le Syndrome de l’Intestin Irritable (SII) mais aussi organiques comme les Maladies Inflammatoires Chroniques de l’Intestin (MICI). De plus, ces pathologies malgré leurs différences dans la sévérité et les signes cliniques, partagent un certain nombre de caractéristiques physiopathologiques communes, comme la rupture de l’intégrité de la barrière intestinale associée à une rupture de tolérance vis-à-vis des antigènes luminaux et plus particulièrement contre le microbiote intestinal. L’objectif de ce travail a été d’évaluer les effets d’un Stress de Séparation Maternelle (SSM) sur l’homéostasie intestinale et la réponse immunitaire vis-à-vis du microbiote commensal chez la souris jeune adulte. Nos résultats ont mis en évidence un dimorphisme sexuel dans ce modèle. Chez les souris mâles jeunes adultes, le SSM diminue les fonctions de la barrière intestinale associé à une altération de la réponse humorale et cellulaire au niveau systémique vis-à-vis du microbiote commensal, ainsi qu’à un défaut des cellules présentatrices d’antigènes, conduisant à une inflammation de bas grade malgré un profil proinflammatoire des lymphocytes T. Chez les souris femelles jeunes adultes, le SSM altère la fonctionnalité des cellules de Paneth associée à surpopulation bactérienne intestinale, responsable de la sensibilité viscérale en réponse à une distension colorectale et une réponse humorale systémique dirigée contre le microbiote commensal. Nous avons mis en évidence une empreinte du SSM chez le jeune adulte, capable d'induire des modifications profondes de l’homéostasie intestinale ainsi que de la réponse immunitaire systémique contre le microbiote intestinal. Le SSM altère l'homéostasie intestinale et reproduit des caractéristiques communes aux pathologies digestives à savoir une rupture de barrière associée à une réponse immunitaire contre le microbiote sans symptômes majeurs. Ce travail a permis d'identifier la survenue d'événements stressants pendant la petite enfance comme un facteur environnemental important capable d'altérer l'homéostasie intestinale chez des individus sains et qui pourrait contribuer au déclenchement de pathologies intestinales chez des individus génétiquement prédisposés. / Perinatal period is characterized by an immature intestinal barrier particularly permeable to luminal antigens and as such highly vulnerable to environmental factors like toxins, infections or stressful events. The appropriate maturation of intestinal barrier leads to intestinal homeostasis and tolerance toward luminal contents. Early life stressful events are associated with the development and/or maintenance of functional gastrointestinal disorders like Irritable Bowel Syndrome (IBS) or organic one like Inflammatory Bowel Diseases (IBD). These pathologies are highly different in term of etiology and clinical severity however they share common features like alteration in intestinal barrier associated with an abnormal immune response toward luminal contents especially commensal microbiota. Our aim was to evaluate the consequences of maternal separation (MS) on intestinal homeostasis, host-microbiota relationship and the humoral and cellular response at adulthood. Due to sexual dimorphism in this model, the results are presented separately for male and female. In young adult male mice, MS decreases intestinal barrier functions associated with an alteration of systemic humoral and cellular response toward commensal microbiota. Moreover, a defect of antigen presenting cells in spleen leads to systemic low grade inflammation despite a pro-inflammatory profile of T cell. In young adult female mice, MS alters the functionality of Paneth cells associated with an intestinal bacterial overgrowth, leading to visceral sensitivity and systemic humoral response toward commensal microbiota. We highlighted that MS has long lasting adverse effects on intestinal homeostasis and systemic immune response toward commensal microbiota in young adult. MS impairs intestinal homeostasis in healthy individuals and might contribute to trigger intestinal pathologies in susceptible persons.

<p>Karakterizacija&nbsp; ekstrakata&nbsp; plodnih&nbsp; tela&nbsp; devet&nbsp;autohtonih&nbsp; vrsta&nbsp; gljiva&nbsp; pripadnika&nbsp; razdela&nbsp;Basidiomycota&nbsp; obuhvatila&nbsp; je&nbsp; preliminarno&nbsp;ispitivanje&nbsp; hemijskog&nbsp; sastava&nbsp; i&nbsp; ispitivanje biolo&scaron;ke&nbsp; aktivnosti.&nbsp; Primenom&nbsp; HPLC‐MS/MS&nbsp; i&nbsp;GC-FID tehnike &nbsp;detektovan&nbsp;je&nbsp;i&nbsp;određen&nbsp;sadržaj&nbsp;fenolnih&nbsp;jedinjenja&nbsp;i&nbsp;metil&nbsp;estara&nbsp;masnih&nbsp;kiselina.&nbsp;<br />Primenom&nbsp; tehnike&nbsp; elektroforeze&nbsp; na&nbsp; mikročipu&nbsp;(&quot;LAB‐ON‐A‐CHIP&quot;&nbsp; kapilarnom&nbsp; elektroforezom)&nbsp;izvr&scaron;ena&nbsp;je&nbsp;karakterizacija&nbsp;proteinskih&nbsp;frakcija&nbsp;u&nbsp;odabranim&nbsp; vrstama&nbsp; gljiva.&nbsp; Antioksidantna&nbsp;aktivnost&nbsp; (sposobnost&nbsp; neutralizacije&nbsp; slobodnih &nbsp;radikala, &nbsp;redukcioni&nbsp;potencijal),&nbsp;sadržaj&nbsp;ukupnih&nbsp;fenola&nbsp; i&nbsp; flavonoida&nbsp; vodenih&nbsp; i&nbsp; etanolnih&nbsp;<br />ekstrakata&nbsp; ispitan&nbsp; je&nbsp; primenom&nbsp;spektrofotometrijskih&nbsp; metoda.&nbsp; Antimikrobna&nbsp;<br />aktivnost&nbsp; metanolnog&nbsp; i&nbsp; heksanskog&nbsp; ekstrakta&nbsp;ispitana&nbsp; je&nbsp; na&nbsp; Gram‐pozitivnim&nbsp; i&nbsp; Gram&ndash;negativnim bakterijskim&nbsp; sojevima.&nbsp; Efekat etanolnih&nbsp;i&nbsp;vodenih &nbsp;ekstrakata &nbsp;odabranih&nbsp;vrsta gljiva&nbsp; na&nbsp; rast&nbsp;ćelijske&nbsp; linije&nbsp; MCF7&nbsp; (humani adenokarcinom&nbsp;dojke pozitivan&nbsp;na&nbsp;estrogenske&nbsp;i progesteronske&nbsp;receptore),&nbsp;ispitan&nbsp;je&nbsp;MTT&nbsp;i&nbsp;SRB testom,&nbsp;dok&nbsp;je&nbsp;efekat&nbsp;frakcija&nbsp;ispitan&nbsp;primenom MTT&nbsp; testa.&nbsp; Sumarno,&nbsp; ukupni&nbsp; biopotencijal ispitivanih&nbsp;vrsta&nbsp;na&nbsp;osnovu&nbsp;sprovedenih&nbsp;analiza hemijske&nbsp; karakterizacije&nbsp; i&nbsp; biolo&scaron;kih&nbsp; testova upućuju&nbsp;opravdanost&nbsp;hipoteze&nbsp;da&nbsp;su&nbsp;odabrane vrste&nbsp;potencijalni &nbsp;izvori&nbsp;nutraceutika.&nbsp;Svakako&nbsp;je potrebno&nbsp;i&nbsp;neophodno&nbsp;nastaviti&nbsp;dalja&nbsp;istraživanja ovih&nbsp;vrsta.</p> / <p>Characterization&nbsp;of&nbsp;extracts&nbsp;of&nbsp;fruiting&nbsp;bodies&nbsp;of&nbsp;<br />nine&nbsp;native&nbsp;species&nbsp;of&nbsp;mushrooms&nbsp;members&nbsp;of&nbsp;<br />section&nbsp; Basidiomycota&nbsp; included&nbsp; preliminary&nbsp;<br />testing&nbsp; of&nbsp; chemical&nbsp; composition&nbsp; and&nbsp; biological&nbsp;<br />activity.&nbsp; Using&nbsp; HPLC‐MS&nbsp; /&nbsp; MS&nbsp; and&nbsp; GC‐FID&nbsp;<br />techniques&nbsp;the&nbsp;content&nbsp;of&nbsp;phenolic&nbsp;compounds&nbsp;<br />and&nbsp;fatty&nbsp;acid&nbsp;methyl&nbsp;esters&nbsp;has&nbsp;been&nbsp;detected&nbsp;<br />and&nbsp;determined.&nbsp;By&nbsp;applying&nbsp;the&nbsp;techniques&nbsp;of&nbsp;<br />electrophoresis&nbsp; on&nbsp; microchip&nbsp; (&quot;LAB‐ON‐A‐CHIP&quot;&nbsp;<br />capillary&nbsp; electrophoresis)&nbsp; characterization&nbsp; of&nbsp;<br />protein&nbsp;fractions&nbsp;in&nbsp;selected&nbsp;types&nbsp;of&nbsp;mushrooms&nbsp;<br />has&nbsp;been&nbsp;performed.&nbsp;The&nbsp;antioxidant&nbsp;activity&nbsp;(the&nbsp;<br />ability&nbsp; of&nbsp; neutralizing&nbsp; free&nbsp; radicals,&nbsp; reducing&nbsp;<br />potential),&nbsp; the&nbsp; contents&nbsp; of&nbsp; total&nbsp; phenols&nbsp; and&nbsp;<br />flavonoids&nbsp; of&nbsp; water&nbsp; and&nbsp; ethanol&nbsp; extracts&nbsp; has&nbsp;<br />been&nbsp; tested&nbsp; using&nbsp; the&nbsp; spectrophotometric&nbsp;<br />method.&nbsp; The&nbsp; antimicrobial&nbsp; activity&nbsp; of&nbsp; methanol&nbsp;<br />and&nbsp; hexane&nbsp; extracts&nbsp; was&nbsp; tested&nbsp; against&nbsp; Gram‐<br />positive&nbsp;and&nbsp;Gram‐negative&nbsp;bacterial&nbsp;strains.&nbsp;The&nbsp;<br />effect&nbsp; of&nbsp; ethanol&nbsp; and&nbsp; aqueous&nbsp; extracts&nbsp; of&nbsp;<br />selected&nbsp;fungi&nbsp;species&nbsp;on&nbsp;the&nbsp;growth&nbsp;of&nbsp;the&nbsp;cell&nbsp;<br />line&nbsp; MCF7&nbsp; (human&nbsp; breast&nbsp; adenocarcinoma&nbsp;<br />positive&nbsp; for&nbsp; estrogen&nbsp; and&nbsp; progesterone&nbsp;<br />receptors),&nbsp; has&nbsp; been&nbsp; examined&nbsp; using&nbsp; MTT&nbsp; and&nbsp;<br />SRB&nbsp;assay,&nbsp;while&nbsp;the&nbsp;effect&nbsp;of&nbsp;the&nbsp;fractions&nbsp;has&nbsp;<br />been&nbsp;tested&nbsp;by&nbsp;using&nbsp;the&nbsp;MTT&nbsp;assay.&nbsp;In&nbsp;summary,&nbsp;<br />the&nbsp;overall&nbsp;biological&nbsp;potential&nbsp;of&nbsp;the&nbsp;examined&nbsp;<br />species&nbsp; based&nbsp; on&nbsp; results&nbsp; for&nbsp; chemical&nbsp; and&nbsp;<br />biological&nbsp; characterization&nbsp; tests&nbsp; (anti‐oxidative,&nbsp;<br />antimicrobial&nbsp;and&nbsp;anti‐proliferative)&nbsp;indicate&nbsp;the&nbsp;<br />justifiability&nbsp; of&nbsp; the&nbsp; hypothesis&nbsp; that&nbsp; examined&nbsp;<br />species&nbsp; of&nbsp; mushrooms&nbsp; are&nbsp; potential&nbsp; sources&nbsp; of&nbsp;<br />nutraceuticals.&nbsp; It&nbsp; is&nbsp; certainly&nbsp; needed&nbsp; and&nbsp;<br />necessary&nbsp;to&nbsp;continue&nbsp;further&nbsp;research&nbsp;of&nbsp;these&nbsp;<br />species.</p>

<p>Upotrebom 50% etanola i 50% acetona kao ekstragensa izvr&scaron;ena je ekstrakcija delova ploda kestena: srž ploda, braon spoljna kora ploda, crvena unutra&scaron;nja kora ploda, ceo plod (bez ježevica), kao i drveta: li&scaron;će, resa, ježevice, stara i mlada kora stabla. Ispitivani su pitomi kesten, lovranski marun i kalemljeni italijanski marun. Nakon određivanja prinosa suvog ekstrakta, primenom standardnih spektrofotometrijskih metoda određen je sadržaj fenolnih jedinjenja, flavonoida i kondenzovanih tanina. Iako je primenom 50% acetona kao ekstragensa dobijen veći prinos fenolnih materija, flavonoida i kondenzovanih tanina, za proizvodnju ekstrakata se preporučuje 50% etanol kao ekstragens, jer obezbeđuje sasvim zadovoljavajuće rezultate, a prihvatljiviji je sa aspekta znatno niže toksičnosti.<br />Ekstrakt lista lovranskog maruna i rese pitomog kestena roda 2007. godine &scaron;tite eritrocite od hemolize izazvane H₂O₂.<br />Glavne komponente hidrolizata nakon metanolizacije ekstrakata su dimetil estar dehidrodigalne kiseline i metil estar dilaktona elaginske i valoneinske kiseline. Kvantitativnom LC/MS i HPLC/DAD analizom najveći sadržaj elagitanina je utvrđen za ekstrakt ježevica (170,6 mg/g ekstrakta).<br />Izv&scaron;eno je ispitivanje antioksidativne aktivnosti ekstrakata u odnosu na 1,1-difenil-2-pikrilhidrazil (DPPH), hidroksi (^&bull;OH) i superoksidni anjon (^&bull;O₂^-) radikale. Kapacitet ekstrakata za transformaciju organskih hidrofilnih radikala, ispitan kao sposobnost redukcije spin probe Tempon je najveći za ekstrakt rese pitomog kestena (A = 18,1%), dok ekstrakti lista pitomog kestena i mlade kore drveta ispoljavaju slabu aktivnost. Ispitivanje protektivnog delovanja ekstrakata u odnosu na UV zračenje određeno je kao sposobnost uklanjanja ^&bull;OH i ^&bull;O₂^- radikala nastalih nakon zračenja. Ekstrakti koji ispoljavaju pozitivne, ali relativno niske RI vrednosti za proizvodnju obe vrste radikala ^&bull;OH i ^&bull;O₂^- su spoljna braon kora lovranskog maruna, resa sa kalemljenog italijanskog maruna i list lovranskog maruna. Negativne RI vrednosti dobijene za ostale ekstrakte ukazuju na prooksidativnu aktivnost u vodenom rastvoru izloženom UV zračenju. Ekstrakti rese, lista i ježevica ispoljavaju aktivnost u cilju preveniranja/otklanjanja lipidne peroksidacije membrane eritrocita.<br />Ispitivanjem<em> in vitro</em> antioksidativne aktivnosti primenom MTT testa je utvrđeno da ekstrakti rese i ježevica pitomog kestena i lista lovranskog maruna imaju izuzetno visoku antioksidativnu aktivnost u ćeliji. Naročito je povoljna činjenica da deluju u niskim koncentracijama (0,02 mg/ml). Antimikrobna aktivnost ekstrakata <em>C. sativa </em>određena je u odnosu na (G+) bakterije: <em>S. aureus, S. lutea, B. cereus, L. lactis ssp. lactis, M. pyrogenes var. albus</em>, kao i na (G-) bakterije: <em>P. mirabilis i S. typhimurium</em>. Značajnu antimikrobnu aktivnost daju ekstrakti kore drveta, ježevica i spoljne braon kore ploda. Ekstrakti srži ploda i celog ploda nisu ispoljili antimikrobnu aktivnost. Postoji značajna i jako značajna korelacija između antimikrobne aktivnosti ekstrakata, kao i antimikrobne i antioksidativne aktivnosti u odnosu na superoksid anjon radikale. Ekstrakti lista, ježevica, spoljne braon i unutra&scaron;nje crvene kore ploda, kao i kore drveta <em>C. sativa</em> Mill. dobijeni primenom 50% etanola kao ekstragensa predstavljaju značajan izvor komponenata sa farmakolo&scaron;kim delovanjem u cilju smanjenja nivoa oksidativnog stresa, poseduju visok kapacitet sprečavanja lipidne peroksidacije, deluju u pravcu preveniranja/otklanjanja lipidne peroksidacije i za&scaron;tite membrane eritrocita, imaju visoku <em>in vitro</em> antioksidativnu aktivnost, a ispoljavaju i značajnu antimikrobnu aktivnost.</p> / <p>Parts of chestnut such as: seeds (without spiny burs), peeled chestnut, red internal seed coat and brown seed coat, as well as parts of the trees: leaf, catkin, spiny burs, young and old chestnut bark have been extracted by 50% ethanol and 50% acetone as an extragent. Three cultivars of <em>Castanea sativa</em> Mill.: sweet chestnut, Lovran&#39;s marrone and grafted Italian marrone were examined. After determination of the yield of dry extract, the content of total phenolic compounds, flavonoids and condensed tannins are determined by application of standard spectrophotometrics methods. Although, the highest content of total phenolics, flavonoids and condensed tannins are obtained by 50% acetone as extragents, for production of extracts 50% ethanol is more suitable, regards much lower toxicity.<br />Extracts of the leaf of Lovran&#39;s marrone and catkin of sweet chestnut native in 2007, protect erythrocytes from hemolysis provoked by H₂O₂.<br />Dehydrodigallic acid dimethyl ester, ellagic acid and valoneic acid dilactone methyl ester are the main compounds in all hydrolysates after methanolisation. The highest content of ellagitannin was detected in extract of spiny burs (170.6 mg/g extract), by application of quantitative LC/MS and HPLC/DAD analysis.<br />The examination of antioxidant activity of 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl (^&bull;OH) and superoxide (^&bull;O2^-) radicals have been done. Capacity of extracts for removal of organic, hydrophilic radicals, exanimate as potential of reduction of spin probe Tempon is highest in extract of catkin of sweet chestnut (A = 18.1%), while extracts of catkin, leaf and spiny burs almost have no antioxidative activity. The evaluation of UV-protective activity of extracts is determinated as capacity for removal of ^&bull;OH and O₂^- radicals generated after irradiation. Extracts which showed positive, but relative low RI values for production of both radical species, OH and ^&bull;O₂ radicals, are brown seed coat of Lovran&#39;s marrone, catkin of grafted Italian marrone and leaf of Lovran&#39;s marrone. Negative RI values obtained for other extracts show that these have prooxidative activity in aqueous solution exposed to UV radiation. Extracts of catkin, leaf and spiny burs expressed activity to prevent/remove lipid peroxidation in the membrane of erythrocytes.<br />Examination of antioxidant activity <em>in vitro</em> by application of MTT test have been detected especially high antioxidant activity of extracts of catkin, spiny bur of sweet chestnut and leaf of Lovran&rsquo;s marrone in the cell. Particularly is favorable that extracts acting in low concentration (0.02 mg/ml). Antimicrobial activity of extracts of <em>C. sativa</em> was determinated against Gram-positive bacteria:<em> S. aureus, S. lutea, B. cereus, L. lactis ssp. lactis, M. pyrogenes var. albus</em>, as well as Gram-negative bacteria: <em>P. mirabilis and S. typhimurium</em>. The significant antimicrobial activity shows extracts of bark, spiny burs and brown seed coat. Extracts of peeled chestnut and seeds didn&rsquo;t show any antimicrobial activity. The very significant and significant correlation existed between antimicrobial activity of extracts, as well as antimicrobial activity and scavenging of^&bull;O2^- radical. Extracts of leaf, spiny burs, brown seed coat and red internal seed coat, as well as bark of <em>C. sativa</em> Mill. produced by 50% ethanol as extragent represent important resource of components with pharmacological activity in reducing level of oxidative stress, possess high activity to prevent/remove lipid peroxidation and protection of the membrane of erythrocytes, have high<em> in vitro</em> antioxidant activity, and also express significant antimicrobial activity.</p>

<p>U disertaciji je ostvarena sineza amida i oksazolina žučnih kiselina, kao i njihovih alkil i alkilidenskih derivata polazeći od holne kiseline. Ipitano je pona&scaron;anje različitih okso derivata žučnih kiselina u uslovima Grignard-ove i Wittig-ove reakcije. Ispitana je biolo&scaron;ka aktivnost odabranih sintetizovanih jedinjenja</p> / <p>Synhesis of bile acid amide and oxazoline derivatives, and their alkyl and alkylidene derivatives was accomplished starting from cholic acid. Also, chemical behavior of different bile acid oxo derivatives in Grignard and Wittig reaction was investigated. Biological activity&nbsp; of selected synthesized compounds was evaluated.</p>

<p>U ovoj doktorskoj disertaciji je radi fizičko-hemijske karakterizacije i ispitivanja potencijalne<br />biolo&scaron;ke aktivnosti sintetisano&nbsp; 40&nbsp; derivata mono(tio)karbohidrazona. Strukture dobijenih jedinjenja su potvrđene&nbsp; NMR&nbsp; i&nbsp; FT&ndash;IR&nbsp; spektroskopijom,&nbsp; elementalnom&nbsp; i&nbsp; rendgenskom&nbsp; strukturnom&nbsp; analizom. Uticaj&nbsp; rastvarača&nbsp; i&nbsp; prisutnog&nbsp; supstituenta&nbsp; na&nbsp; položaje apsorpcionih&nbsp; maksimuma&nbsp; ispitani&nbsp; su&nbsp; po principima LSER i LFER metoda. Teorijski proračuni za sintetisana jedinjenja izvedeni su&nbsp; primenom TD&ndash;DFT&nbsp; metode.&nbsp; Jonizacione&nbsp; konstante&nbsp; derivata&nbsp; mono(tio)karbohidrazona&nbsp; su&nbsp; određene potenciometrijskim&nbsp; titracijama.&nbsp; Antioksidativna&nbsp; aktivnost&nbsp; je&nbsp; određena&nbsp; primenom&nbsp; tri&nbsp; testa:&nbsp; DPPH, ABTS&nbsp; i&nbsp; FRAP&nbsp; testa.&nbsp; Antimikrobna&nbsp; aktivnost&nbsp; je&nbsp; ispitana&nbsp; mikrodilucionom&nbsp; metodom&nbsp; na&nbsp; dva&nbsp; soja bakterija:&nbsp; Staphylococcus&nbsp; aureus&nbsp; i&nbsp; Escherichia&nbsp; coli.&nbsp; Za&nbsp; jedinjenja&nbsp; koja&nbsp; su&nbsp; pokazali&nbsp; antimikrobnu aktivnost je&nbsp; ispitan procenat citotoksičnosti.&nbsp; Sveukupan&nbsp; zaključak o sličnosti i razlikama sintetisanih derivata izveden je na osnovu multivarijatnih metoda klaster analize i analize glavnih komponenti.</p> / <p>In this doctoral dissertation, 40 mono(thio)carbohydrazone derivatives were synthesized for the purpose of physico-chemical characterization and examination of potential biological activity. The structures of the obtained compounds were confirmed by NMR and FT &ndash;IR spectroscopy, elemental analysis&nbsp; and&nbsp; X-ray&nbsp; structural&nbsp; analysis.&nbsp; The&nbsp; influence&nbsp; of&nbsp; solvent&nbsp; and&nbsp; substituent&nbsp; present&nbsp; on&nbsp; the positions&nbsp; of&nbsp; absorption maxima&nbsp; was&nbsp; examined&nbsp; according&nbsp; to&nbsp; the&nbsp; principles&nbsp; of&nbsp; LSER&nbsp; and&nbsp; LFER methods.&nbsp; Theoretical&nbsp; calculations&nbsp; for&nbsp; the&nbsp; synthesized&nbsp; compounds&nbsp; were&nbsp; performed&nbsp; by&nbsp; the&nbsp; TD&ndash;DFT method.&nbsp; The&nbsp; acid&nbsp; constants&nbsp; of&nbsp; mono(thio)carbohydrazone&nbsp; derivatives&nbsp; were&nbsp; determined&nbsp; by potentiometric&nbsp; titrations.&nbsp; Antioxidant&nbsp; activity&nbsp; was&nbsp; determined&nbsp; using&nbsp; three&nbsp; tests:&nbsp; DPPH,&nbsp; ABTS&nbsp; and FRAP test. Antimicrobial activity was examined by microdilution method on two strains of bacteria: Staphylococcus&nbsp; aureus&nbsp; and&nbsp; Escherichia&nbsp; coli.&nbsp; The&nbsp; percentage&nbsp; of&nbsp; cytotoxicity&nbsp; was&nbsp; examined&nbsp; for compounds&nbsp; that&nbsp; showed&nbsp; antimicrobial&nbsp; activity.&nbsp; Overall&nbsp; conclusion&nbsp; about&nbsp; the&nbsp; similarities&nbsp; and differences of the synthesized&nbsp; derivatives was studied by multivariate&nbsp; methods of cluster analysis and analysis of principal components.</p>

Tesis por compendio / [ES] El uso masivo de plásticos y su impacto medioambiental, así como la necesidad de mejorar la conservación de los alimentos, hace necesario el desarrollo de alternativas biodegradables para el envasado activo de alimentos. La presente Tesis Doctoral, se ha centrado en el desarrollo de películas activas, basados en polímeros biodegradables con propiedades complementarias, con el fin de obtener materiales con propiedades mecánicas y de barrera adecuadas para cumplir con los requisitos del envasado alimentario. Para ello, se consideraron el poli (vinil alcohol) (PVA) y el poli (ácido láctico) PLA. Asimismo, el carvacrol y los ácidos ferúlico y cinámico fueron elegidos como compuestos activos naturales. Se estudió la encapsulación del carvacrol en liposomas de diferentes tipos de lecitina para mejorar su retención durante la etapa de formación del film. La lecitina de soja enriquecida en fosfatidilcolina fue la más eficaz para mantener la estabilidad de la emulsión de carvacrol, resultando en films con mayor retención. La incorporación de carvacrol (encapsulado o no), modificó ligeramente la microestructura y las propiedades físicas de los films, así como la cristalinidad y el comportamiento térmico del polímero. El PVA totalmente hidrolizado proporcionó films con un mejor desempeño mecánico y capacidad de barrera al oxígeno, pero con menor retención de carvacrol. Por el contrario, el PVA parcialmente hidrolizado dio lugar a films más homogéneos, con mayor contenido de carvacrol. Mientras que los grupos acetilos protegieron al polímero de la termodegradación, permitiendo la obtención de films mediante las técnicas habituales del procesado industrial. Por lo tanto, el PVA parcialmente hidrolizado tiene un gran potencial para la producción de films activos, con más posibilidades de procesamiento que el PVA totalmente hidrolizado. Se incorporaron también ácido cinámico y ferúlico, con menor impacto sensorial que el del carvacrol, al PVA parcial y totalmente hidrolizado, mediante casting de las soluciones acuosas con glicerol para mejorar la solubilidad de los ácidos. Los films de PVA plastificados con glicerol mostraron una menor capacidad de barrera que los no plastificados. La incorporación del ácido ferúlico promovió mayores cambios en las propiedades de los films que el ácido cinámico, debido a un efecto de reticulación, que promovió la cristalinidad, la rigidez y la capacidad de barrera del material. También se obtuvieron películas plastificadas a base de PVA parcialmente hidrolizado con ácidos fenólicos, mediante mezclado en fundido y moldeo por compresión. Los films termoprocesados fueron menos extensibles y resistentes a la rotura y más permeables al oxígeno y al vapor de agua. Estudios in vitro demostraron que los films con ácidos fenólicos obtenidos por casting o termoprocesado presentaron actividad antioxidante y antimicrobiana, especialmente con ácido ferúlico. Con el objetivo de cumplir los requisitos de envasado de alimentos, se obtuvieron films tricapa por termocompresión, compuestos por una capa central de PVA, cargada con compuestos activos (carvacrol o ácido ferúlico), y dos capas externas de PLA. La termocompresión fue eficaz para la adhesión interlaminar. Las multicapas mostraron un rendimiento mecánico próximo al de los films de PLA y una elevada capacidad de barrera al oxígeno y al vapor de agua. Todos los laminados con compuestos activos incorporados a la capa de PVA fueron eficaces para controlar el crecimiento microbiano en filetes de carne de vacuno envasada. Por tanto, el laminado de films de PVA y PLA parcialmente hidrolizado representa una estrategia adecuada para obtener materiales de envasado con propiedades funcionales más próximas a las de algunos plásticos convencionales utilizados en el envasado alimentario. Asimismo, estos materiales con carvacrol o ácido ferúlico incorporados podrían utilizarse para el envasado de alimentos altamente susceptibles a procesos oxidativos o de degradación microbiológica para alargar su vida útil. / [CA] L'ús massiu de plàstics i el seu impacte mediambiental, així com la necessitat de millorar la conservació dels aliments, fa necessari el desenvolupament d'alternatives biodegradables per a l'envasament actiu d'aliments. La present Tesi Doctoral, s'ha centrat en el desenvolupament de pel·lícules actives, basats en polímers biodegradables amb propietats complementàries, amb la finalitat d'obtindre materials amb propietats mecàniques i de barrera adequades per a complir amb els requisits de l'envasament alimentari. Per a això, es van considerar el poli (vinil alcohol) (PVA) i el poli (àcid làctic) PLA. Així mateix, es van triar el carvacrol i els àcids ferúlic i cinàmic, com a compostos actius. Es va estudiar l'encapsulació del carvacrol en liposomes de diferents tipus de lecitina per a millorar la seua retenció durant l'etapa de formació del film. La lecitina de soja enriquida en fosfatidilcolina va ser la més eficaç per a mantindre l'estabilitat de l'emulsió de carvacrol, resultant en films amb major retenció. La incorporació de carvacrol (encapsulat o no), va modificar lleugerament la microestructura i les propietats físiques dels films, així com la cristalinitat i el comportament tèrmic del polímer. El PVA totalment hidrolitzat va proporcionar films amb un millor exercisc mecànic i capacitat de barrera a l'oxigen, però amb menor retenció de carvacrol. Per contra, el PVA parcialment hidrolitzat va donar lloc a films més homogenis, amb major contingut de carvacrol. Mentre que els grups acetils van protegir el polímer de la termodegradació, permetent l'obtenció de films mitjançant les tècniques habituals del processament industrial. Per tant, el PVA parcialment hidrolitzat té un gran potencial per a la producció de films actius, amb una més àmplia gamma de possibilitats de processament que el PVA totalment hidrolitzat. Es van incorporar també àcid cinàmic i ferúlic, amb menor impacte sensorial que el del carvacrol, al PVA parcial i totalment hidrolitzat, mitjançant càsting de les solucions aquoses amb glicerol per a millorar la solubilitat dels àcids. Els films de PVA plastificats amb glicerol van mostrar una menor capacitat de barrera que els no plastificats. La incorporació de l'àcid ferúlic va promoure majors canvis en les propietats dels films que l'àcid cinàmic, a causa d'un efecte de reticulació, que va promoure la cristalinitad, la rigidesa i la capacitat de barrera del material. També es van obtindre pel·lícules plastificades a base de PVA parcialment hidrolitzat amb àcids fenòlics, mitjançant barrejat en fos i emotlament per compressió. Els films termoprocessats van ser menys extensibles i resistents al trencament i més permeables a l'oxigen i al vapor d'aigua. Estudis in vitro van demostrar que els films amb àcids fenòlics obtinguts per càsting o termoprocessat van presentar activitat antioxidant i antimicrobiana, especialment amb àcid ferúlic. Amb l'objectiu de complir els requisits d'envasament d'aliments, es van obtindre films tricapa per termocompressió, compostos per una capa central de *PVA, carregada amb compostos actius (carvacrol o àcid ferúlic), i dues capes externes de PLA. La termocompressió va ser eficaç per a l'adhesió interlaminar. Les multicapes van mostrar un rendiment mecànic pròxim al dels films de PLA i una elevada capacitat de barrera a l'oxigen i al vapor d'aigua. Tots els laminatges amb compostos actius incorporats a la capa de PVA van ser eficaces per a controlar el creixement microbià en filets de carn de boví envasada. Per tant, el laminatge de films de PVA i PLA parcialment hidrolitzat representa una estratègia adequada per a obtindre materials d'envasament amb propietats funcionals més pròximes a les d'alguns plàstics sintètics utilitzats en l'envasament alimentari. Així mateix, aquests materials amb carvacrol o àcid ferúlic incorporats podrien utilitzar-se per a l'envasament d'aliments altament susceptibles a processos oxidatius o de degradació microbiològica per a allargar la seua vida útil. / [EN] The massive use of synthetic plastics and their environmental impact, as well as the need of improving food preservation, makes necessary the development of biodegradable alternatives for food active packaging. The use of active compounds from natural origin to obtain this kind of materials is a new trend based of the consumer demand for safer and healthier products. The present Doctoral Thesis, has been focused on the development of active packaging films, based on biodegradable polymers with complementary properties, in order to obtain materials with adequate mechanical and barrier properties to meet food packaging requirements. To this end, poly (vinyl alcohol) (PVA) and poly (lactic acid) (PLA) were considered. Likewise, carvacrol and ferulic and cinnamic acid, were chosen as active natural compounds. The liposomal encapsulation of carvacrol, using different kinds of lecithin was evaluated in order to improve the carvacrol retention in the polymer matrix during the film formation step. Soy lecithin enriched in phosphatidylcholine was the most effective at maintaining the stability of the carvacrol emulsion, leading to its highest retention in the PVA films. Incorporation of carvacrol (lecithin encapsulated or not) slightly modified the film microstructure and physical properties and polymer crystallinity and thermal behaviour. The fully hydrolysed PVA provided films with a better mechanical performance and oxygen barrier capacity, but with less carvacrol retention. In contrast, partially hydrolysed PVA gave rise to more homogenous films with a higher carvacrol content. Acetyl groups protect the polymer for thermodegradation, allowing the film production using the common industrial thermoplastic processing techniques. Therefore, partially hydrolysed PVA has a great potential for the production of active films, with a wider range of processing possibilities than fully hydrolysed PVA. Cinnamic and ferulic acids, with lower potential sensory impact than of carvacrol, were also incorporated into partially and fully hydrolysed PVA by casting of aqueous solutions containing glycerol to improve the acid solubility. Glycerol plasticized PVA films exhibited poorer barrier capacity than those non-plasticized. The incorporation of ferulic acid promoted greater changes in the film properties than cinnamic acid, due to a crosslinking effect, which promoted crystallinity, stiffness and barrier capacity of the material. Glycerol plasticized films based on partially hydrolysed PVA with phenolic acids were also obtained by melt blending and compression moulding. Termoprocessed films were less stretchable and resistant to break and more permeable to oxygen and water vapor. In vitro studies demonstrated that cast and termoprocessed films containing phenolic acids exhibited antioxidant and antimicrobial activity, especially with ferulic acid. Three-layer films composed of a central layer of PVA loaded with active compounds (carvacrol lecithin encapsulated or not, or ferulic acid) and two external PLA layers were obtained by thermo-compression, aimed to meet food packaging requirements. Thermocompression was effective for the interlaminar adhesion. Multilayers exhibited mechanical performance close to the PLA sheets and high oxygen and water vapour barrier capacity. All laminates with active compounds incorporated into the PVA sheet were effective at controlling microbial growth in packaged beef meet. Therefore, lamination of partially hydrolysed PVA and PLA films represents a successful strategy to obtain packaging materials with functional properties closer to those of some conventional plastics commonly used in food packaging. Likewise, these materials incorporating carvacrol or ferulic acid could be used for packaging of foods highly susceptible to oxidative or microbiological degradative process to extend their shelf life. / The authors acknowledge the financial support of the Agencia Estatal de Investigacion, Ministerio de Economía y Competitividad (MINECO) of Spain through the projects AGL2016-76699-R and PID2019-105207RB-100. Author Johana Andrade thanks the Departamento de Nariño-Colombia y la Fundación CEIBA for the doctoral grant. The authors also thank the services rendered by the Electron Microscopy Service of the UPV. / Andrade Chapal, JC. (2021). Biodegradable materials based on poly(vinyl alcohol) (PVA) and poly (lactic acid) (PLA) with antioxidant and antimicrobial activity for food packaging applications [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/175566 / Compendio

Rapid development of bacterial resistance and multiresitance to conventional antibiotics has resulted in an intensive search for alternative antimicrobial agents. Antimicrobial peptides (AMPs) belong to promising anti-infective candidates since they do not development bacterial resistance. They kill microbes by disturbing or permeabilizing the cytoplasmic membrane, or may target putative key intracellular compartments. Their advantages include fast action and selectivity between prokaryotic and eukaryotic cells. We have isolated several novel AMPs from the venom of wild bees: halictines (HAL-1 and HAL-2) from Halictus sexcinctus, lasiocepsin (Las) from Lasioglossum laticeps and macropin (MAC-1) from Macropis fulvipes. They are active against Gram-positive and Gram- negative bacteria and against yeast Candida albicans. While halictines and macropin have moderate hemolytic activity, Las shows no hemolytic activity. A novel AMP was isolated also from the mucus of Xiphydria camelus. This AMP belongs to the category of insect defensins. It contains 55 amino acid residues, three disulphide bridges and its C-terminus is amidated. CD and NMR studies of HAL-1, HAL-2 and MAC-1 revealed propensity to form amphipathic α-helical structure in the presence of sodium dodecyl sulfate or trifluoroethanol. For the...

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 2017-07-24T19:22:05Z (GMT). No. of bitstreams: 1 FELIPE GUTIERREZ REYES.pdf: 2973646 bytes, checksum: 0e8c702449c97c861e9f0314e3b69e2f (MD5) Previous issue date: 2016-02-23 / Objectives: This study evaluated the effect of addition of copper nanoparticles at different concentrations into a simplified etch-and-rinse (ER) adhesive system (Ambar [AM]) on antimicrobial activity (AMA), degree of conversion of adhesives discs (DC-d) and dentin-resin interface (DC-i), the ultimate tensile strength (UTS), 28-day cumulative water sorption (WS), solubility (So) and copper release (CR) as well as immediate (IM) and 1-year (1Y) resin–dentine bond strength (μTBS) and nanoleakage (NL). Methods: Seven experimental adhesive systems were formulated according to the addition of copper nanoparticles (0 [control], 0.0075, 0.015, 0.06, 0.1, 0.5 and 1%) in AM adhesive system. We tested the antimicrobial activity of synthesized adhesives against Streptococcus mutans using agar diffusion assays. For DC-d, specimens were constructed and tested after 24 h for FTIR and micro-Raman spectroscopy. For UTS, specimens were tested after 24 h and 28 days. For WS and So, after specimens build-up, they were stored in water and the properties measured for 28 days. For CR, specimens were stored in 2% nitric acid solution and the properties were measured for 28 days. The occlusal enamel of thirty-five molars was removed and adhesives were applied to dentine surface after 37% phosphoric acid etching. After composite resin build-ups, specimens were longitudinally sectioned to obtain resin–dentine bonded sticks (0.8 mm2). Specimens were tested in tension at 0.5 mm/min in IM or 1Y (μTBS). For NL, 2 bonded sticks from each tooth were prepared and analyzed under SEM in IM or 1Y. For DC-i, 2 bonded sticks were prepared and analyzed under micro-Raman spectroscopy. The μTBS and NL data of each adhesive were subjected to two-way repeated measures ANOVA. For UTS, WS, So, copper release, DC-d and DC-i data of each adhesive were subjected to a one-way ANOVA. Tukey’s post hoc test was used for pair-wise comparisons (α = 0.05). Results: The addition of copper nanoparticles provided antimicrobial properties to the adhesives at all concentrations (p < 0,05), and did not influence UTS, WS and SO (p > 0.05). Higher CR was observed in adhesives with higher concentration of copper nanoparticles (p < 0.05). The addition of 1% of copper nanoparticles decreased the DC-d and DC-i significantly. After 1Y, significant reductions of μTBS and increases of NL were observed in the control group (p < 0.05). Reductions of μTBS and increase of NL over time were not observed for copper-containing adhesives.Conclusions: The addition of copper nanoparticles in concentrations up to 0.5% in the simplified ER Ambar adhesive system may be an alternative to provide antimicrobial properties, increase the long-term stability of resin–dentine interfaces, 12 without reducing adhesives’ mechanical properties evaluated. / Objetivos: Este estudo avaliou o efeito da adição de nanopartículas de cobre, em diferentes concentrações, em um sistema adesivo convencional (CON) simplificado Ambar [FGM] sobre a atividade antimicrobiana (AAM), grau de conversão de corpos de prova de adesivos (GC-c) e na interface de união a dentina (GC-i), resistência máxima à tração (RT), sorção cumulativa de água (SO), solubilidade (SB) e liberação de cobre (LC) durante 28 dias, bem como a resistência de união (RU) e nanoinfiltração (NI) na interface de união à dentina nos tempos imediatos (IM) e após 1 ano (1A). Material e métodos: Sete sistemas adesivos experimentais foram formulados de acordo com a adição de nanopartículas de cobre (0 [controle], 0,0075, 0,015, 0,06, 0,1, 0,5 e 1%) no sistema adesivo Ambar. Foi testada a atividade antimicrobiana dos adesivos formulados contra Streptococcus mutans por meio de ensaios de difusão em ágar. Para GC-c, as amostras foram construídas e testadas após 24 h por meio de espectrofotometria no infravermelho com Transformação de Fourier (FTIR) e espectroscopia de micro-Raman. Para RT, as amostras foram testadas depois de 24 horas e 28 dias. Para SO e SB, depois de confeccionados os espécimes, eles foram armazenados em água e as propriedades medidas por 28 dias. Para LC, as amostras foram armazenadas em solução de ácido nítrico a 2% e as propriedades medidas durante 28 dias. O esmalte oclusal de trinta e cinco molares foi removido e os adesivos foram aplicados na superfície da dentina depois de condicionamento com ácido fosfórico a 37%. Depois de restaurações de resina composta, os espécimes foram seccionados longitudinalmente para se obter espécimes (palitos) de resina-dentina (0,8 mm2). Os espécimes foram testados em tensão a 0,5 mm/min no IM ou 1A (RU). Para NI, 2 espécimes de cada dente foram preparadas e analisadas em MEV e testados no IM ou 1A. Para GC-i, 2 espécimes foram preparados e analisados sob espectroscopia de micro-Raman. Os dados do RT foram analisados por uma análise de variância de 2 fatores. Para AAM, SO, SB, e liberação de cobre, os dados foram analisados por análise de variância de fator único, e para RU e NI os dados foram submetidos a análise de variância de 2 fatores para medidas repetidas. Também foi feito o teste post hoc de Tukey para múltiplas comparações. A significância estatística foi predefinida em α = 0,05.Resultados: A adição de nanopartículas de cobre adicionou propriedades antimicrobianas aos adesivos em todas as concentrações (p < 0,05), e não influenciou a RT, SO e SB (p > 0,05). LC elevadas foram observadas em adesivos com maior concentração de nanopartículas de cobre (p < 0,05). A adição de 1% de 10 nanopartículas de cobre diminuiu significativamente o GC-c e GC-i. Após 1 ano, decréscimos significativos de RU e aumentos de NI foram observados no grupo controle (p < 0,05). Decréscimos significativos de RU e aumentos de NI não foram observados nos adesivos contendo cobre (p > 0,05). Conclusões: A adição de nanopartículas de cobre em concentrações de até 0,5% no sistema adesivo convencional simplificado Ambar pode ser uma alternativa para adicionar atividade antimicrobiana e aumentar a estabilidade a longo prazo das interfaces resina-dentina, sem comprometer as propriedades mecânicas dos adesivos.

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