- 4 - ABSTRACT During the revision of epiphytic species of the Lecanora subfusca group in the Czech Republic, nine taxa have been recorded. L. cinereofusca and L. exspersa are reported for the first time from the country. L. circumborealis has been excluded from the list of Czech lichens. L. cinereofusca and L. horiza could be considered as extinct. L. rugosella and L. subrugosa are regarded as extreme morpholocial forms from L. chlarotera and L. argentata. These morphotypes corelate with ecological conditions: substrate (nutrient enriched bark) and habitat (eutrophisation effect). In case of L. subrugosa, this speculation was confirmed by molecular data (ITS rDNA sequences). Several new secondary metabolites have been discovered during the detailed research of chemical lichen compounds. These substances belonging to terpenoids are taxonomically important and very helpful for distinguishing single species. The abundance of L. allophana and L. chlarotera has decreased during last decades. Contrarily, L. pulicaris expanded slightly. The main reasons of changes in distribution are air pollution and acidification of substrates as the impact of acid rains. The rate of substrate acidification has been shown on example of L. pulicaris. In the past, this lichen predominated on acid-barked porophytes. Nowadays, it...

Les extraits n-hexaniques respectivement des feuilles d’Hopea odorata Roxb. et du bois de Dipterocarpus costatus Gaertn.f. ont été sélectionnés sur la base d’un criblage biologique préliminaire. Ces extraits présentent des activités cytotoxiques significatives sur les lignées MCF-7 (cancer du sein) et NCI-H187 (cancer du poumon non à petite cellule). De plus, l’extrait du bois de D. costatus inhibe la croissance de la souche K1 de Plasmodium falciparum. L’étude phytochimique de l’extrait n-hexanique des feuilles d’Hopea odorata a permis d’isoler 19 composés terpéniques dont 16 triterpènes des séries lupane (n=8), (3,4-seco)cycloartane (n=4), friedélane (n=2) et oléanane (n=2). Parmi les lupanes, l’acide 3,30-dioxolup-20(29)-èn-28-oïque a été isolé pour la première fois d’une source naturelle. L’évaluation de l’effet de ces lupanes sur la croissance de quatre lignées cancéreuses humaines (PC3, MDA-MB-231, HT-29 and HCT116) a permis d’identifier des composés actifs et d’apporter des éléments de relations structure-activité tels que l’influence du degré d’oxydation des positions C-3, C-28 et C-30 sur la cytotoxicité. Parmi les cycloartanes, deux nouveaux 3,4-secocycloartanes ont été identifiés. Il s’agit de deux esters d’acide gras saturés et de l’acide (24S, 25S, 26)-trihydroxy-3,4-secocycloart-4(29)-èn-3-oïque. Par ailleurs, l’étude phytochimique de l’extrait n-hexanique du bois de Dipterocarpus costatus a permis d’isoler 31 terpènes dont 12 nouveaux triterpènes, parmi lesquels 5 norlupanes, 3 dammaranes, 2 nordammaranes et 2 secodammaranes. Les activités biologiques de l’ensemble des composés isolés, ont été évaluées sur la croissance des lignées cancéreuses humaines précédemment citées et de muscle squelettique de rat L-6, ainsi que sur la croissance de la souche K1 du Plasmodium falciparum. En particulier, le norlupane 36 possédant une fonction endoperoxyde, montre une forte activité antiplasmodiale associée à une faible cytotoxicité. / Preliminary study of the extracts from Hopea odorata leaves and Dipterocarpus costatus wood showed that the hexane extracts of both plants exhibited cytotoxicity against breast cancer (MCF-7) and small cell lung cancer (NCI-H187), in conjunction with potent antimalarial (against Plasmodium falciparum K1 strain) activity of D.costatus wood hexane extract. Phytochemical studies of H. odorata leaves hexane extract led to the isolation of 16 terpenoids in the series lupane (n = 8), (3,4-seco) cycloartane (n = 4), friedelane (n = 2) and oleanane (n = 2). Among lupanes, 3,30-dioxolup-20(29)-en-28-oic acid was isolated for the first time from a natural source. Cytotoxicity of lupanes triterpenes against four human cancer cell lines (PC3, MDA-MB-231, HT-29 and HCT116) was evaluated and showed the structure activity relationship related to the oxidation degree at position 3, 28 and 30. Among cycloartanes two new 3,4-seco-cycloartanes were identified. They are saturated fatty acid esters of (24S,25S,26)-trihydroxy-3,4-seco-cycloart-4(29)-en-3-oic acid. Isolation of D. costatus wood hexane extract discovered 31 terpenoids of which 12 triterpenes are new, including 5 norlupanes, 3 dammaranes, 2 nordammaranes and 2 secodammaranes. The biological activities of all isolated compounds were evaluated through cytotoxicity against human cancer cell lines mentioned above and rat myoblast-derived cells L-6, as well as antimalarial activity against Plasmodium falciparum K1 strain. Interestingly, the norlupane 36, possessing an endoperoxide group, showed a strong antiplasmodial activity associated with low cytotoxicity.

<p>Karakterizacija metanolnih ekstrakata i etarskih ulja iglica i &scaron;i&scaron;arki osam vrsta samoniklih kleka (rod Juniperus L.) obuhvatala je fitohemijski skrining i ispitivanje antioksidantne, antiinflamatorne i antimikrobne aktivnosti. Primenom LC-MS/MS i GC-MS tehnike detektovan i odreĎen je sadržaj fenolnih i terpenoidnih sekundarnih biomolekula. Antioksidantna aktivnost ekstrakata i etarskih ulja (sposobnost neutralizacije slobodnih radikala, redukcioni potencijal i inhibicija lipidne peroksidacije) ispitana je primenom spektrofotometrijskih metoda. U cilju odreĎivanja antiinflamatornog potencijala, primenjena je ex vivo metoda za praćenje aktivnosti trombocitne ciklooksigenaze-1 i 12-lipoksigenaze. Antimikrobna aktivnost etarskih ulja odreĎena je na &scaron;est odabranih bakterijskih sojeva. Ispitani ekstrakti i etarska ulja pokazali su značajnu biolo&scaron;ku aktivnost.</p> / <p> Characterization of methanol extracts and essential oils of eight Juniperus L. species included phytochemical screening and evaluation of antioxidant, anti-inflammatory and antimicrobial activity. The presence and content of phenolics and terpenoids was confirmed by LC-MS/MS and GC-MS technique. Antioxidant activity of extracts and essential oils (radical scavenger capacity, reduction potential and inhibition of lipid peroxidation) was examined by spectrophotometric methods. With the intention to evaluate anti-inflammatory activity, an ex vivo method was applied to measure activity of platelet cyclooxygenase-1 and 12-lipoxygenase. Antimicrobial activity was evaluated according to six bacterial strains. Examined extracts and essential oils showed noticeable biological activity.</p>

The presente work describes the results obtained from the phytochemical studies bioguided of crude extracts of the bark of A. salzmannii, a species belonging to the Annonaceae family, front to tests of antioxidant, antimicrobial, and insecticide activities against larvae of the mosquito Aedes aegypti. From the hexane extract were isolated five compounds, been three sesquiterpenes (caryophyllene oxide, spathulenol, and selin-11-en-4a-ol); and two steroids (stigmast-4-en-3-one and - sitosterol). From the chloroform alkaloidal fraction of the methanolic extract were isolated seven alkaloids: liriodenine, 10-methoxyliriodenine, anonaine, asimilobine, reticuline, cleistopholine, and a mixture of alkaloids anonaine and xylopine. The assay of antioxidant activity revealed that the methanol extract (3583.02 μmol of TE g-1) and its fractions, chloroform alkaloidal fraction (5291.77 μmol of TE g-1) and chloroform neutral fraction (2047.54 μmol of TE g-1) showed significant antioxidant activity, mainly the chloroform alkaloidal fraction. Among the alkaloids isolated and tested, asimilobine was the most active of them with antioxidant activity of 2.09 TE relative. For antimicrobial assay, the best results were observed for methanol extract and chloroform alkaloidal fraction against Staphylococcus aureus (ATCC14458), Escherichia coli (ATCC 10799), Candida tropicalis (ATCC 157), and C. dubliniensis (ATCC 777). Among the alkaloids isolated and tested, all showed significant results with MIC values between 25-100 mg.mL-1, mainly the alkaloids anonaine, asimilobine, and liriodenine against Staphylococcus aureus penicilinase- (8-), S. epidermidis (ATCC 12228), S. epidermidis (6ep), and Candida dubliniensis (ATCC 778157). To the test of larvicidal activity, only the hexane extract and the fractions of methanol extract showed activity below 700 mg.mL-1. The caryophyllene oxide showed activity of 167 mg.mL-1. The results obtained confirm that A. salzmannii is a typical species of the Annonaceae family, and a promising source of biologically active compounds. / O presente trabalho descreve os resultados obtidos a partir do estudo fitoquímico biomonitorado dos extratos brutos das cascas de Annona salzmannii, uma espécie pertencente a família Annonaceae, frente a testes de atividades antioxidante, antimicrobiana e larvicida contra o Aedes aegypti. Do extrato hexânico foram isoladas cinco substâncias, sendo três sesquiterpenos (óxido de cariofileno, espatulenol e selin-11-en-4a-ol); e dois esteróides (estigmast-4-en-3-ona e - sitosterol). Da fração clorofórmica alcalóidica, proveninente do extrato metanólico, isolou-se sete alcalóides: liriodenina, 10-metoxiliriodenina, anonaina, asimilobina, reticulina, cleistofolina e uma mistura dos alcalóides anonaina e xylopina. O ensaio de atividade antioxidante revelou que o extrato metanólico (3583,02 μmol de TE g-1) e suas respectivas frações, fração clorofórmica alcalóidica (5291,77 μmol de TE g-1) e fração clorofórmica neutra (2047,54 μmol de TE g-1) apresentaram expressiva atividade antioxidante, com destaque para a fração clorofórmica alcaloídica. Dentre os alcalóides isolados e testados, asimilobina foi o mais ativo com atividade antioxidante de 2,09 TE relativo. Para o ensaio de atividade antimicrobiana, os melhores resultados foram observados para o extrato metanólico e a fração clorofórmica alcaloídica contra Staphylococcus aureus (ATCC14458), Escherichia coli (ATCC 10799), Candida tropicalis (ATCC 157) e C. dubliniensis (ATCC 777). Dentre os alcalóides isolados e testados, todos apresentaram resultados significativos com valores de CIM entre 25-100 mg.mL-1, com destaque para os alcalóides anonaina, asimilobina e liriodenina frente a Staphylococcus aureus penicilinase-(8-), S. epidermidis (ATCC 12228), S. epidermidis (6ep) e Candida dubliniensis (ATCC 778157). Para o teste de atividade larvicida, apenas o extrato hexânico e as frações provenientes do extrato metanólico apresentaram atividade abaixo de 700 mg.mL-1. O óxido de cariofileno apresentou atividade de 167 mg.mL-1. Os resultados obtidos confirmam que A. salzmannii é quimicamente uma espécie típica da família Annonaceae, além de uma fonte promissora de substâncias biologicamente ativas.

Submitted by isabela.moljf@hotmail.com (isabela.moljf@hotmail.com) on 2017-06-21T14:43:48Z No. of bitstreams: 1 viniciuscariusdesouza.pdf: 3785363 bytes, checksum: 7063d6c5f5cef353643903ad5f125c48 (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2017-08-07T19:09:52Z (GMT) No. of bitstreams: 1 viniciuscariusdesouza.pdf: 3785363 bytes, checksum: 7063d6c5f5cef353643903ad5f125c48 (MD5) / Made available in DSpace on 2017-08-07T19:09:52Z (GMT). No. of bitstreams: 1 viniciuscariusdesouza.pdf: 3785363 bytes, checksum: 7063d6c5f5cef353643903ad5f125c48 (MD5) Previous issue date: 2016-03-03 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico / FAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas Gerais / Lippia alba, popularmente conhecida por erva-cidreira, é uma espécie vegetal amplamente distribuída pelas Américas e encontrada praticamente em todo o território brasileiro. Esta espécie possui importante uso na medicina tradicional para o tratamento de cólicas, indigestão, náuseas, espasmos, diarreia, disenteria, doenças respiratórias, problemas hepáticos e no tratamento de sífilis e gonorreia. As folhas de L. alba, as quais são preparadas sob a forma de infusão ou decocção e ingeridas por via oral, produzem um óleo essencial rico em moléculas iso-prenóides denominadas terpenóides. Estes compostos não são apenas de interesse farmacológico, mas também industrial já que são usados na confecção de fragrâncias. A composição dos óleos essenciais pode variar em função de diferentes fatores abióticos e genotípicos, como por exemplo nível de ploidia. Neste contexto, os objetivos deste trabalho foram caracterizar o transcriptoma de folha da espécie L. alba e buscar sequencias putativas de enzimas envolvidas na produção de metabólitos secundários. O transcriptoma foi sequenciado pela plataforma Miseq (Illumina) com bibliotecas pairedend de 300 bp. O sequenciamento resultou em um total de 47.498.310 reads paired-end (23.749.155 reads para cada end sequenciado) de 35-308 bp, compreendendo 12.148.327.567 nucleotídeos (-12 Gb). A montagem de novo dos transcritos foi processada a partir do software Trinity que gerou 193.532 transcritos, sendo 128.209 unigenes, com o valor de N50 igual a 1.187 bp. Um total de 86.122 ORF (Open Read Frame) foi obtido e a seguir submetido ao algoritmo de alinhamentos BlastP, o qual encontrou 75.533 sequências com referência no banco de dados NR (Non-Redundant) de proteínas. Aproxima-damente, 78,4% dessas sequências foram anotadas funcionalmente a partir do pipeline utiliza-do pelo software Blast2GO. As análises das sequências anotadas revelaram prováveis enzimas para síntese de terpenóides como geraniol e linalol/nerolidol. Para validação da montagem e anotação, foram realizados ensaios de qPCR para amplificação de sequências de 13 genes para controles endógenos e 4 genes de terpeno sintases. Os resultados obtidos aqui corroboram outros estudos de transcriptoma de espécies não modelo usando tecnologias de sequenciamento de alto-desempenho. / Lippia alba, popularly known as erva-cidreira, is a widely distributed specie in Americas and it is found throughout Brazil. This specie has important using in popular medicine for cramp-ing, indigestion, nausea, diarrhea, dysentery, respiratory diseases, liver disorders treatment and infectious diseases such as syphilis and gonorrhea. The leaves of L. alba, which are pre-pared by infusion or decoction and orally ingested, producing an essential oil rich in terpene compounds. These compounds are of pharmacological and industrial interest, due to their use in fragrance preparation. Interestingly, the composition of essential oils change according to different abiotic factors and genetic variations such as ploidy level. In this context, the aims of this work were to characterize the transcriptome of leaves of L. alba (linalool chemotype) and to search putative enzymes sequences involved in production of secondary metabolites. The transcriptome was sequenced by Miseq platform (Illumina) running pair-end libraries 300 bp. The sequencing resulted in 47,498,310 reads (23,749,155 reads for each end sequenced) of 35-308 bp, comprising 12,148,327,567 nucleotides (-12 Gb). The de novo assembly of tran-scripts was processed by Trinity software and generated 193,532 transcripts, in 128,209 uni-genes, with N50 equal to 1,187 bp. 86,122 ORFs (Open Read Frame) were obtained and sub-mitted to BlastP algorithm, finding 75,533 sequences included in NR (Non-Redundant) pro-tein database. Approximately 78.4% of these sequences were functionally annotated using Blast2Go pipeline. Analysis of annotated sequences revealed putative enzymes for synthesis of terpenoids such as geraniol and linalool/nerolidol. For assembly and annotation validation, qPCR assay were realized by amplification of 13 endogenous control genes and 4 terpene synthases genes. The results found here corroborate transcriptome studies in non-model or-ganisms using high-performance sequencing technologies.

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