Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Markéta Marešová Supervisor: Ing. Petra Matoušková, Ph.D. Title of diploma thesis: Effect of alpha-humulene on adhesion of breast cancer cell line MDA-MB-231 α-humulene is a sesquiterpene contained in the essential oil of Chinese Bayberry (Myrica rubra), which has various biological effects. The aim of this thesis was to study the cell adhesion of tumor cell line MDA-MB-231, and the effects of α-humulene on expression of cell adhesion molecules. Furthermore, the cytotoxic effect of α-humulene on this cell line was verified. Effect of α-humulene on cell proliferation was evaluated by neutral red uptake test (NRU test) and by xCelligence system. Cell adhesion was also continuously monitored by xCelligence. Expression ganges of cell adhesion molecules upon α-humulene treatment were determined by Western blotting and by quantitative polymerase chain reaction on the protein and mRNA levels, respectively. The proliferation of the cells was significantly affected by α-humulene after 24 hour treatment (IC50 13.6 µg/ml). α-humulene alone caused slightly increased cell adhesion. Adhesion molecules EpCAM and β-cathenin were almost unaffected. Level of ICAM1 mRNA was increased after 12 hours and...

Made available in DSpace on 2014-06-11T19:32:39Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-05-06Bitstream added on 2014-06-13T20:04:01Z : No. of bitstreams: 1 montanarijunior_i_dr_botfca.pdf: 1131496 bytes, checksum: 0a7962bc58e62cb2a1d6049912fcf8d6 (MD5) / Este trabalho avaliou agronômica e quimicamente 37 progênies de meios irmãos de Cordia verbenacea DC., espécie nativa de importância medicinal por causa de sua ação antiinflamatória. O objetivo desta pesquisa foi o de identificar progênies e indivíduos de C. verbenacea com boas características agrícolas e químicas, iniciando assim o processo de domesticação da espécie, visando promover seu cultivo, de modo a oferecer uma nova opção agrícola aos produtores rurais, contribuindo para a preservação da espécie e para utilização dos seus benefícios terapêuticos. Através de um ensaio agrícola em blocos casualizados, com 4 repetições, foram avaliadas o comportamento de progênies de meios irmãos em três colheitas. As características analisadas foram: a produção de biomassa verde, de folhas secas, o índice de colheita, a produção de óleo essencial e a produção de alfa humuleno. A partir dos dados obtidos foi estimada a variabilidade disponível para o melhoramento, a herdabilidade das características analisadas, o progresso esperado com a seleção, a eficiência do método de seleção empregado e as correlações genéticas, ambientais e fenotípicas entre as características analisadas. Concluiu-se que as progênies possuíam ampla variação genética e que esta variação poderia ser explorada para fins de melhoramento. As herdabilidades calculadas para massa de folhas secas, porcentagem de óleo essencial e massa de óleo essencial foram de 0,52, 0,69 e 0,78 respectivamente, considerando-se as três colheitas. Calculando-se os ganhos por seleção, concluiu-se que podem ser esperados ganhos expressivos selecionando-se as melhores plantas dentro das melhores progênies. O método de seleção entre e dentro de progênies de meios irmãos empregado neste trabalho mostrou-se significantemente mais eficiente que os métodos de seleção entre progênies... / This dissertation evaluated agronomically and chemically 37 progenies of half-sib families of Cordia verbenacea DC., a native medicinal species important for its anti-inflammatory action. The objective of this research was to identify C. verbenacea individuals with good agricultural and chemical characteristics to start a domestication process, aiming at promoting cultivation and offering a new agricultural option to rural producers, thus contributing to preserve the species and to the use of its therapeutic properties. An agricultural experimental design of randomized blocks with four repetitions evaluated the behavior of half-sib families in three harvests. The characteristics analyzed were: fresh biomass and dry leaves production, harvest index, essential oil production, and alpha humulene production. The available variability for breeding, the heritability of the analyzed characteristics, the progress expected from selection, the efficiency of the chosen selection method, and the genetic, environmental, and phenotypic correlations between the characteristics analyzed were estimated from the collected data. It was concluded that the progenies had a broad genetic variability that could be explored for breeding. The heritabilities calculated for dried leaf biomass, essential oil percentage, mass of essential oil were, respectively, 52%, 0,69% and 0,78%, considering the three harvests. On estimating the gain from selection, the conclusion was that significant gain can be expected from the selection of the best plants between and within the best progenies. The selection method between and within half-sib families employed in this research proved to be considerably more efficient than methods such as selection between progenies, and mass and stratified mass selection. The correlations study shows that oil and alpha humulene production is closely related to biomass production, demonstrating... (Complete abstract click electronic access below)

FundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgico / Este estudo foi realizado com o objetivo de encontrar novas fontes vegetais do antiinflamatÃrio natural α-humuleno que jà à utilizado na elaboraÃÃo de fitomedicamento comercial. Para alcanÃar este propÃsito, realizou-se a extraÃÃo atravÃs de duas tÃcnicas (arraste a vapor e hidrodestilaÃÃo) dos Ãleos essenciais de sete espÃcies: Persea americana Mill, Eclipta prostrata L, Ocimum gratissimum L, Ambrosia artemisiaefolia L, Plectranthus ornatus Cold, Lantana camara L e Aloysia virgata (R. et P.) Juss. Os extratos obtidos foram analisados atravÃs de cromatografia gÃs-lÃquÃdo acoplada a espectrÃmetro de massas (CG/EM) e com detector de ionizaÃÃo de chama (CG-DIC). AlÃm do α-humuleno, foram identificados majoritariamente β-cariofileno (34,62%) em Persea americana Mill por hidrodestilaÃÃo, cedr-8-eno (20,20% e 15,39%, respectivamente por arraste à vapor e hidrodestilaÃÃo) em Ambrosia artemisiaefolia L; β-cariofileno (51,53% e 39,73%, respectivamente por arraste à vapor e por hidrodestilaÃÃo) nas folhas de Eclipta prostata L, de Lantana camara L (31,43% e 25,50%) e de Plectranthus ornatus Cold (18,33% e 27,08%). O Ãleo de Ocimum gratissimum Mill apresentou principalmente eugenol (45,42% e 40,96%, respectivamente por arraste à vapor e por hidrodestilaÃÃo). Em Aloysia virgata Juss. por arraste a vapor, β-cariofileno (16,30%) e por hidrodestilaÃÃo, epÃxido de cariofileno (29,05%) foram identificados. O α-humuleno foi isolado parcialmente atravÃs de CLAE com detector de UV-VIS. A avaliaÃÃo das atividades antiinflamatÃria, antioxidante, larvicida e anticolinesterÃsica dos extratos volÃteis das espÃcies em estudo e dos extratos hexÃnico e etanÃlico das folhas da Aloysia virgata (R. et P.) Juss. foram realizadas. Os resultados indicaram os Ãleos essenciais de Ocimum gratissimum L, Ambrosia artemÃsiaefolia L e Lantana camara L, como agentes antiinflamatÃrios naturais. / This study was aimed at finding new sources of natural anti-inflammatory α-humulene, which is already used in the preparation of commercial phytodrug. For this purpose, the extraction was performed by two techniques (steam distillation and hydrodistillation) of essential oils of seven species: Persea americana Mill, Eclipta prostrata L, Ocimum gratissimum L, Ambrosia artemisiaefolia L, Plectranthus ornatus Cold, Lantana camara L and Aloysia virgata (R. et P.) Juss. The extracts were analyzed by GC / MS) and GC-FID. In addition to the α-humulene were identified mainly β-caryophyllene (34.62%) in Persea americana Mill by hydrodistillation, cedar-8-ene (20.20% and 15.39%, respectively, by steam distillation and hydrodistillation) in Ambrosia artemisiaefolia L, β-caryophyllene (51.53% and 39.73% respectively by steam distillation and hydrodistillation) in the leaves of Eclipta prostata L, of Lantana camara L (31.43% and 25.50%) and of Plectranthus ornatus Cold (18.33% and 27.08%). The oil of Ocimum gratissimum Mill presented mainly eugenol (45.42% and 40.96% respectively by steam distillation and hydrodistillation). In Aloysia virgata Juss. by steam distillation, β-caryophyllene (16.30%) and by hydrodistillation, caryophyllene epoxide (29.05%) were identified. The α-humulene was partially isolated using HPLC with UV-VIS detector. The evaluation of anti-inflammatory activity, antioxidant and larvicidal anticholinesterase volatile extracts of the species under study and the hexane and ethanol extracts of leaves of Aloysia virgata (R. et P.) Juss. were performed. The results indicated the essential oils of Ocimum gratissimum L, Ambrosia artemÃsiaefolia L and Lantana camara L, as natural anti-inflammatory agents.

Accurate measurements of ammonia, nitric acid and formic acid are important for achieving a complete understanding of their atmospheric role. Models and measurements of formic acid in the atmosphere continue to show disagreements. Also, the contributions of NMHCs and reactive nitrogen (HNO3 and NH3) to organic and inorganic aerosol formation are important to quantify as gaps in the knowledge of atmospheric aerosols are a source of uncertainty in climate science. In this thesis, concentrations of ammonia were measured in the atmosphere and the production of formic acid from the ozonolysis of isoprene was measured in the EXTRA (EXTreme RAnge) chamber. Both gases were studied using chemical ionisation mass spectrometry (CIMS). The kinetics of the reactions of sesquiterpenes and terminal alkenes with ozone were studied in theEXTRA chamber using the relative rates technique and GC-FID. The ozonolysis rate coefficients of a homologous series of terminal alkenes were measured at elevated temperatures and found to be invariant with the carbon number. This led to the conclusion that previous measurements of these rate coefficients were subject to experimental artefacts. The elevated temperature protocol was employed to study the ozonolysis of sesquiterpenes, leading to revisions in ko3 for β-caryophyllene and α-humulene of 3 orders of magnitude. It was thus concluded that ozonolysis only accounts for 9-15% of sesquiterpeneoxidation in the atmosphere. A field intercomparison of CIMS for measuring ammonia was conducted wherein CIMS was found to perform well alongside instruments of comparable time response and limits of detection. This thesis also characterised inlet materials used in atmospheric measurements in the first systematic study on the uptake onto inlet walls in a flow tube system coupled to CIMS. It was found that PFA is the preferable material for atmospheric measurements, both for its kinetic qualities and its ready availability and ease of use. Finally, CIMS was used to measure the yield of formic acid from isoprene ozonolysis as a function of relative humidity. Formic acid yield was found to increase between 0-40% RH to a maximum of 0.18. Using the measured formic acid yields in a global chemistry model leads to an estimate that formic acid production from isoprene ozonolysis is ~9.5 Tg yr-1.

Orientador: Norberto da Silva / Banca: Lin Chau Ming / Banca: Eduardo Pagani / Banca: Joaquim Adelino de Azevedo Filho / Banca: Walter José Siqueira / Resumo: Este trabalho avaliou agronômica e quimicamente 37 progênies de meios irmãos de Cordia verbenacea DC., espécie nativa de importância medicinal por causa de sua ação antiinflamatória. O objetivo desta pesquisa foi o de identificar progênies e indivíduos de C. verbenacea com boas características agrícolas e químicas, iniciando assim o processo de domesticação da espécie, visando promover seu cultivo, de modo a oferecer uma nova opção agrícola aos produtores rurais, contribuindo para a preservação da espécie e para utilização dos seus benefícios terapêuticos. Através de um ensaio agrícola em blocos casualizados, com 4 repetições, foram avaliadas o comportamento de progênies de meios irmãos em três colheitas. As características analisadas foram: a produção de biomassa verde, de folhas secas, o índice de colheita, a produção de óleo essencial e a produção de alfa humuleno. A partir dos dados obtidos foi estimada a variabilidade disponível para o melhoramento, a herdabilidade das características analisadas, o progresso esperado com a seleção, a eficiência do método de seleção empregado e as correlações genéticas, ambientais e fenotípicas entre as características analisadas. Concluiu-se que as progênies possuíam ampla variação genética e que esta variação poderia ser explorada para fins de melhoramento. As herdabilidades calculadas para massa de folhas secas, porcentagem de óleo essencial e massa de óleo essencial foram de 0,52, 0,69 e 0,78 respectivamente, considerando-se as três colheitas. Calculando-se os ganhos por seleção, concluiu-se que podem ser esperados ganhos expressivos selecionando-se as melhores plantas dentro das melhores progênies. O método de seleção entre e dentro de progênies de meios irmãos empregado neste trabalho mostrou-se significantemente mais eficiente que os métodos de seleção entre progênies... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: This dissertation evaluated agronomically and chemically 37 progenies of half-sib families of Cordia verbenacea DC., a native medicinal species important for its anti-inflammatory action. The objective of this research was to identify C. verbenacea individuals with good agricultural and chemical characteristics to start a domestication process, aiming at promoting cultivation and offering a new agricultural option to rural producers, thus contributing to preserve the species and to the use of its therapeutic properties. An agricultural experimental design of randomized blocks with four repetitions evaluated the behavior of half-sib families in three harvests. The characteristics analyzed were: fresh biomass and dry leaves production, harvest index, essential oil production, and alpha humulene production. The available variability for breeding, the heritability of the analyzed characteristics, the progress expected from selection, the efficiency of the chosen selection method, and the genetic, environmental, and phenotypic correlations between the characteristics analyzed were estimated from the collected data. It was concluded that the progenies had a broad genetic variability that could be explored for breeding. The heritabilities calculated for dried leaf biomass, essential oil percentage, mass of essential oil were, respectively, 52%, 0,69% and 0,78%, considering the three harvests. On estimating the gain from selection, the conclusion was that significant gain can be expected from the selection of the best plants between and within the best progenies. The selection method between and within half-sib families employed in this research proved to be considerably more efficient than methods such as selection between progenies, and mass and stratified mass selection. The correlations study shows that oil and alpha humulene production is closely related to biomass production, demonstrating... (Complete abstract click electronic access below) / Doutor