O café é a mais importante mercadoria no comércio agrícola internacional, representando uma significante fonte de renda para diversos países, incluindo o Brasil. A formação de gemas florais é um importante processo fisiológico em plantas superiores e o período de florescimento é influenciado pelas condições ambientais. Em Coffea arabica L. estudos tem demonstrado que os tecidos dos ramos e do caule são locais de armazenamento de carboidratos de reserva. Entretanto, muito pouco é conhecido sobre a relação existente entre o acúmulo de amido nos tecidos de reserva e o desenvolvimento reprodutivo nesta espécie. O objetivo deste presente estudo foi investigar a variação sazonal do amido nos tecidos de armazenamento de Coffea arabica L cultivar Catuaí vermelho e a relação com a iniciação e desenvolvimento de gemas florais. Este trabalho foi conduzido em condições de campo, utilizando plantas adultas crescendo na Fazenda Areão, o centro experimental da Escola Superior de Agricultura \"Luiz de Queiroz\"-ESALQ. Ramos laterais (plagiotrópicos), caule principal e raízes laterais foram coletados para determinar os locais de armazenamento nas plantas, bem como para caracterizar a variação sazonal de amido em diferentes estágios fenológicos da planta. Lâminas de secções transversais foram preparadas e então submetidas às observações histológicas. Ramos plagiotrópicos que apresentaram gemas florais e ramos sem gemas foram igualmente analisados para determinar a presença ou ausência de amido nos tecidos. Adicionalmente, o amido foi quantificado em tais ramos para determinar a relação entre o nível de carboidratos e o desenvolvimento de gemas. Os resultados indicaram que os ramos laterais, caule principal e raízes são potencialmente locais de armazenamento de reserva. As gemas florais foram observadas em ramos contendo amido acumulado nos tecidos, sugerindo que o desenvolvimento de gemas no cafeeiro arábico é influenciado pela presença deste polissacarídeo. A concentração média de amido nos ramos contendo gemas foi de 42,85 mg.g-1 de matéria seca ou de 9,10 mg.g-1 para ramos sem gemas, indicando a influência dos carboidratos de reserva sobre o ciclo reprodutivo em Coffea arabica. Ramos contendo poucos frutos demonstraram amido nos tecidos de armazenamento e botões florais desenvolvidos, em oposição aos ramos com muitos frutos, mostrando uma clara relação entre tecidos fonte-dreno e a fase reprodutiva no cafeeiro. / Coffee is the most important commodity in the international agricultural trade, representing a significant source of income to several countries, including Brazil. Flower bud formation is one important physiological process for higher plants, and flowering time is influenced by environmental conditions. In Coffea arabica L. studies have demonstrated that tissues of branches and stems are storage sites for reserve carbohydrates. However, little is known about the relationship between starch accumulation and reproductive development in coffee species. The general objective of the current study was to investigate the seasonal variation of starch in the storage tissues of Coffea arabica L. cultivar Red Catuaí and its relation with flower bud initiation and development. This work was carried out in field conditions, using adult plants growing in Areão Farm, the experimental center of Escola Superior de Agricultura \"Luiz de Queiroz\"-ESALQ. Biological samples of plagiotropic branches, main stem, and lateral roots were collected to determinate the storage sites in plant as well as to characterized the seasonal variations of starch in the different phenological stages of plants. Slides of transversal sections of samples were prepared for histological observation. Plagiotropic branches that showed floral buds and branches without buds were even analyzed to determine the presence or absence of starch in tissues. Additionally, starch was quantified in branches to determinate the relation between carbohydrate concentration and bud development. The results indicated that lateral branches, main stem, and roots were potentially reserves sites. The flower buds were only observed in those branches with higher concentrations of starch, as showed by histological preps. This observation suggested that floral bud development in arabica coffe is influenced by the presence of starch reserve. The mean concentration of starch was 42,85 mg.g-1 or 9,10 mg.g-1 dry weight in branches possessing floral buds was to branches without floral buds, respectively. Branches having few fruits showed starch in storage tissues and larger floral buds, opposed to branches with several fruits, showing a clear relationship between source-sink and reproductive phase in coffee plants.

A própolis é uma substância resinosa coletada pelas abelhas de diversas partes das plantas, como brotos, botões florais e exudados resinosos, conhecida por do ácido cinâmico e ácido benzócio, foram ainda encontrados o ácido pimárico, várias atividades biológicas como antimicrobiana, anti-inflamatória, antiproliferativa e antioxidante. Sua composição química depende de vários fatores, como a localização geográfica, vegetação e clima. A própolis brasileira com certificação de produto orgânico, proveniente do sul do Brasil, foi coletada e avaliada em diferentes estações do ano. Essa própolis é produzida em florestas nativas e áreas de reflorestamento, locais livres de contaminação por insumos agrícolas, metais pesados e poluição industrial. O objetivo deste trabalho foi avaliar a composição química e atividade antioxidante, considerando a variação sazonal das estações de verão, outono e primavera. Foram coletadas ao todo 78 amostras provenientes de 14 apiários distintos. O teor de compostos fenólicos totais e flavonoides foram feitos por métodos colorimétricos, a atividade antioxidante foi avaliada pelos métodos de sequestro dos radicais livres DPPH•, ABTS+, sendo a capacidade antioxidante contra os radicais peroxila, determinado pelo método ORAC. O perfil químico do extrato de etanólico da própolis (EEPO) foi avaliado por cromatografia de camada delgada em fase reversa (CCDAE-FR), varredura no ultravioleta-visível (UV-vis) e cromatografia gasosa acoplada com espectrometria de massas (CGEM). A composição de voláteis foi analisada por CG-EM. Os teores de flavonoides variaram de não detectado até 4,76 mg.g-1 e de 6,80 a 72,55 mg.g-1 para fenólicos totais. Em relação à atividade antioxidante, a variação encontrada foi de 1,01 a 384,60 mg Trolox.g-1 para ABTS+, de 4,50 a 148,10 µmol Trolox.g-1 para DPPH• e de 0,20 a 1,25 µmol Trolox.g-1 de amostra para o ORAC. Em relação às estações, o verão apresentou maior teor de flavonoides (p=6%) e o outono, maior teor de DPPH• (p=7%). Com base no perfil químico pela técnica de CCDAE, foi possível classificar as 78 amostras em sete variantes distintas. Dentre os compostos presentes e derivados dos ácidos cinâmico e benzóico, analisados pela técnica CG/MS, identificou-se os compostos ácido pimárico, norolean-12 ene, alfa bisabolol e metil comate A. Também foram identificados oito compostos voláteis em grande quantidade como α-pineno (54,77%), β- pineno (14,83%), α-limoneno (3,78%), β- mirceno (9,29%), ?-candineno (2,11%), γ -muroleno (1,86%), β-felandreno (4,79%) e α-selineno (8,40%). Uma correlação estatística foi encontrada entre o teor de fenólicos totais e o método DPPH•, p=0,78, diferentemente do teor de flavonoides que não apresentou correlação com a atividade antioxidante. Os resultados obtidos demonstraram que a própolis orgânica possui atividade antioxidante, embora apresente baixos teores de flavonoides. Não existe correlação entre os flavonoides e atividade antioxidante. Os compostos fenólicos e a atividade antioxidante foram influenciados pela sazonalidade, sendo o outono, a estação que apresentou teores maiores de fenólicos e atividade antioxidante. / Propolis is a resinous substance collected from plant buds, flowers, and exudates by Apis mellifera bees widely known for its biological activities such as antiviral, antimicrobial, anti-inflammatory, antiproliferative and antioxidant activities. Its chemical composition depends on many factors as geographic location, vegetation, and climate. A certified Brazilian propolis, originated from South Brazil, was collected and evaluated in different seasons. This kind of propolis is produced in native forest and in reforestation areas, where contamination derived from agricultural inputs, heavy metals, and factory fumes is found. Hence, the aim of this work was to evaluate the chemical composition and the antioxidant activity during seasonal variation in summer, autumn and spring. Seventy-eight samples were collected from 14 different apiaries. The phenolic compound content and flavonoids were performed by colorimetric methods. The antioxidant activity was evaluated by free radical scavenging methods such as DPPH• and ABTS+. The antioxidant capacity against peroxil radicals was assessed by ORAC method. The chemical profiles of the organic propolis ethanolic extracts were evaluated by reversed-phase thin-layer chromatography (RP-TLC), ultraviolet (UV) scanning, and gas chromatography-mass spectrometry (GC-MS). Flavonoid content varied from non - detected to 4.76 mg. g-1 and 6.80 to 72.55 mg.g-1 for phenolics. The antioxidant activity varied from 1.01 to 384.60 mg Trolox.g-1 for ABTS+ and 4.50 to 148.10 µmol Trolox.g-1 for DPPH. Considering the seasonal variation, summer presented the highest compound content for flavonoids (p=6%), meanwhile autumn presented the highest compound content for DPPH• (p=7%). Thus, considering the chemical profile presented by RP-TLC, seven different variances of propolis were classified. Compounds such as pimaric acid, norolean-12-ene, alpha bisabolol, and methyl commate A were found by GC-MS technique on these seven variances. Eight volatile compounds were also identified as follow: α-pinene (54,77%), β- pinene (14,83%), α-limonene (3,78%), β-myrcene (9,29%), .-candinene (2,11%), γ - muurolene (1,86%), β-phellandrene (4,79%), and α-selinene (8,40%). In conclusion, the results demonstrated that autumn showed higher antioxidant activity by DPPH• method than those produced in other seasons. A statistical correlation was found between phenolic compound and DPPH•, p=0.78, differing from the flavonoid content which did not demonstrate a correlation with antioxidant activity. The results obtained showed that organic propolis presents antioxidant activity, although its flavonoid content is extremely low. On extent, there was no correlation between flavonoid content and antioxidant activity. Moreover, the seasonal variation showed its influence on phenolic compounds as well as on antioxidant activity, in which autumn presented the highest phenolic content and antioxidant activity.

The thermal environment exerts a strong influence on many aspectsof squamate life history. Environmental temperatures constrain daily activity time, fecundity, growth rate, and also influences hatchling phenotype. Cold nest temperatures can slow the development of eggs deposited in the soil, cause developmental abnormalities or result in mortality of embryos.

Background: Seasonal fluctuations in outdoor temperature have been shown to affect blood pressure in adults. We have asked whether or not there is either a seasonal variation or an influence of outdoor temperature on blood pressure measurements in children and adolescents in middle Europe. Methods: Blood pressure was routinely measured in healthy and sick children and adolescents in outpatient clinics and during hospitalisation in a total of 6714 subjects (3,497 boys/3,237 girls), age 3 to 21 years, with a median of 10.6 years. Results: Cross-sectional analysis showed a significant seasonal variation in blood pressure measurements. The mean increase of systolic and diastolic blood pressure was 4.45/2.42 mmHg during winter. A significant correlation between average outdoor temperatures and systolic blood pressure was found (ρ=-0.074 p<0.001). However, the effect was only detectable at an average temperature below 0° C/32° F and above 10° C/50° F. For each degree Celsius increase in average outdoor temperature, the systolic blood pressure fell by 0.12 mmHg. Conclusion: Blood pressure measurements in children and adolescents, even in a temperate climate, are influenced by temperature and seasonal variation. Considering seasonal variations in blood pressure could be of clinical interest.:Inhaltsverzeichnis......................................................................i Widmung .................................................................................. iii Bibliographische Beschreibung ................................................. iv I. Abkürzungen ......................................................................... i 1. Einführung ........................................................................... 1 1.1. Bedeutung von Blutdruckmessungen im Allgemeinen ................................................................................................. 1 1.2. Bedeutung und Durchführung von Blutdruckmessungen bei Kindern und Jugendlichen ................................................................................................ 2 1.3. Tracking ................................................................................................ 4 1.4. Innere und äußere Einflussfaktoren des Blutdrucks ................................................................................................ 5 1.5. Zweck der Studie ................................................................................................ 7 2. Publikationsmanuskript ................................................................................................ 10 2. 1. Influence of seasonal variation on blood pressure measurements in children, adolescents and young adults ................................................................................................ 10 Abstract .................................................................................. 11 Introduction ................................................................................................ 12 Subjects and methods ................................................................................................ 13 Results .................................................................................... 15 Discussion ................................................................................................ 18 Conclusion ................................................................................................ 21 Acknowledgements ................................................................................................ 22 References............................................................................... 23 Tables ..................................................................................... 27 Figure legends ................................................................................................ 29 Figures ................................................................................... 30 2.2. Blood pressure tracking in children and adolescents ....... 33 Abstract .................................................................................. 34 Inhaltsverzeichnis Subjects and methods ................................................................................................ 35 Results .................................................................................... 39 Discussion ................................................................................................. 42 Conclusion ................................................................................................. 44 Acknowledgements ................................................................................................. 45 References................................................................................ 46 Legends to figures .................................................................................................. 53 Figures ...................................................................................... 55 3. Zusammenfassung der Arbeit ................................................................................................... 62 4. Literaturverzeichnis ............................................................... 68 II. Erklärung über die eigenständige Abfassung der Arbeit ........ ii III. Lebenslauf ............................................................................ iii IV. Danksagung .................................................................................................... iv / Background: High blood pressure is a major risk factor for cardiovascular disease. Blood pressure tracking could help to identify individuals with potential hypertension. Therefore, we have asked whether or not tracking was of predictive value for the development of hypertension in early life. Methods: Blood pressure was routinely measured in 13,261children and adolescents in outpatient clinics as well as during hospitalization. In a sub-analysis 568 individuals with normotensive and elevated blood pressure were compared after three follow-up periods (two, four and six years) and 2,157 normotensive individuals were compared in a paired t-test. Results: The follow-up analysis showed a significant tracking effect. However, the Pearson correlation coefficients of the systolic and diastolic blood pressure SDS decreased over time. Upon the follow-up after six years 35.6% of the children and adolescents with elevated blood pressure values remained in the elevated range group. Of the children within the normotensive blood pressure range 80.4% remained normotensive after six years. Children with normotensive blood pressure showed a stronger tracking than those who had had one hypertensive blood pressure reading. Children with high body weight gain left their SDS track to higher blood pressure values. Conclusion: Blood pressure tracking in children and adolescents is moderate. We conclude that the predictive power of a single hypertensive blood pressure measurement during a single visit is rather small, and thus repetitive measurements across several consecutive visits are necessary.:Inhaltsverzeichnis......................................................................i Widmung .................................................................................. iii Bibliographische Beschreibung ................................................. iv I. Abkürzungen ......................................................................... i 1. Einführung ........................................................................... 1 1.1. Bedeutung von Blutdruckmessungen im Allgemeinen ................................................................................................. 1 1.2. Bedeutung und Durchführung von Blutdruckmessungen bei Kindern und Jugendlichen ................................................................................................ 2 1.3. Tracking ................................................................................................ 4 1.4. Innere und äußere Einflussfaktoren des Blutdrucks ................................................................................................ 5 1.5. Zweck der Studie ................................................................................................ 7 2. Publikationsmanuskript ................................................................................................ 10 2. 1. Influence of seasonal variation on blood pressure measurements in children, adolescents and young adults ................................................................................................ 10 Abstract .................................................................................. 11 Introduction ................................................................................................ 12 Subjects and methods ................................................................................................ 13 Results .................................................................................... 15 Discussion ................................................................................................ 18 Conclusion ................................................................................................ 21 Acknowledgements ................................................................................................ 22 References............................................................................... 23 Tables ..................................................................................... 27 Figure legends ................................................................................................ 29 Figures ................................................................................... 30 2.2. Blood pressure tracking in children and adolescents ....... 33 Abstract .................................................................................. 34 Inhaltsverzeichnis Subjects and methods ................................................................................................ 35 Results .................................................................................... 39 Discussion ................................................................................................. 42 Conclusion ................................................................................................. 44 Acknowledgements ................................................................................................. 45 References................................................................................ 46 Legends to figures .................................................................................................. 53 Figures ...................................................................................... 55 3. Zusammenfassung der Arbeit ................................................................................................... 62 4. Literaturverzeichnis ............................................................... 68 II. Erklärung über die eigenständige Abfassung der Arbeit ........ ii III. Lebenslauf ............................................................................ iii IV. Danksagung .................................................................................................... iv

Os óleos essenciais das folhas e galhos de quatro indivíduos de O. porosa e três de N. megapotamica foram avaliados durante doze meses através das técnica CG-EM e CG-DIC. Os compostos majoritários encontrados nos óleos voláteis das folhas de O. porosa foram durante esse período &#945;-pineno, &#946;-pineno, mirceno, nerolidol e espatulenol, enquanto nos galhos predominavam nerolidol, óxido de cariofileno, &#945;-muurolol, &#946;-eudesmol e o espatulenol, apenas em dois dos quatro indivíduos estudados. Em N. megapotamica, foram observadas diferenças quanti e qualitativas entre os indivíduos para os óleos das folhas e apenas quantitativas para os dos galhos. Nas folhas os componentes majoritários foram &#945;-santaleno, trans-&#945;-bergamoteno, &#946;-santaleno, &#947;-amorfeno, biciclogermacreno e alohimachalol para os indivíduos 1 e 2, enquanto no indivíduo 3 predominavam o &#948;-elemeno e muurola-4, 10 (14)-dien-1-&#946;-ol. Nos galhos desta espécie os componentes majoritários identificados no indivíduo 1 foram &#945;-santaleno, espatulenol, &#945;-muurolol, e 5 iso cedranol; &#945;-santaleno, guaiol, 5 iso cedranol e &#945;-bisabolol, no indivíduo 2; no indivíduo 3 predomina o &#945;-muurolol. A análise dos componentes ao longo dos meses mostrou que os componentes dos óleos estudados não apresentaram uma variação estatisticamente significativa, a variabilidade entre os indivíduos foi maior que aquela observada com a sazonalidade. Os óleos estudados não apresentam atividade fungitóxica para C. sphaerospermum, C. cladosporioides, A. Níger, C. albicans e também não foram ativos frente às bactérias E. coli e P. aeruginosa, somente frente a S. aureus foi observada atividade antibacteriana. / Essential oils extracted from leaves and stems of four individuals of O. porosa and N. megapotamica were evaluated during a period of 12 months using GC/MS and GC/FID techniques. During this period the major compounds found in the volatile oils of O. porosa leaves were &#945;-pinene, &#946;-pinene, myrcene, nerolidol, spathulenol, whereas in stems were nerolidol, caryophyllene oxide, &#945;-muurolol, &#946;-eudesmol and spathulenol, only in two out of the four studied individuals. Among the N. megapotamica specimens qualitative and quantitative differences were observed in the leaf oils but only quantitative in the case of stems. In the leaves of individuals 1 and 2 the major compounds were &#945;-santalene, trans-&#945;- bergamotene, &#946;-santalene, &$947;-amorphene, bicyclogermacrene and alo-himachalol, while for individual 3 &#948;-elemene and muurola-4, 10 (14)-diene-1-&#946;-ol were predominant. In the stems of this species, the major compounds identified were &#945;-santalene, spathulenol, &#945;-muurolol and 5 iso cedranol in the individual 1; &#945;-santalene, guaiol, 5 iso cedranol and &#945;-bisabolol in individual 2; and was the predominant compound &#945;-muurolol in individual 3. The chemical composition of the essential oils, during the analyzed period, did not present a statistically significant variation. Variability among individuals was higher than the one observed with seasonality. The studied oils did not present fungitoxic activity for C. sphaerospermum, C. cladosporioides, A. niger, C. albicans and were also not active against the Gram-negative bacteria E. coli e P. aeruginosa. However, both oils presented some activity against the Gram-positive bacterium S. aureus.

Baccharis dracunculifolia é conhecida por sua interação com insetos, especialmente Apis mellifera L., por possuir ampla diversidade de metabólitos secundários, bem como, por apresentar várias atividades biológicas. Suas folhas são pontuadas por tricomas secretores ricos em metabólitos secundários e também apresentam dutos secretores, estruturas que produzem e armazenam os óleos essenciais. Os metabólitos secundários de B. dracunculifolia são usados pelas abelhas A. mellifera para a produção de própolis ?verde?, que é extremamente valorizada pelas indústrias farmacêuticas, pois apresenta atividades antitumoral, antimicrobiana, antiinflamatória, antiulcerogênica, dentre outras. Considerando estas importantes atividades biológicas, e a necessidade de padronização da própolis, os objetivos do presente trabalho foram: desenvolver metodologia analítica por cromatografia gasosa para quantificar os constituintes majoritários presentes no óleo essencial da planta cultivada; desenvolver e validar metodologia analítica por CLAE capaz de garantir a autenticidade dos extratos hidroalcoólicos de B. dracunculifolia, bem como de própolis verde; determinar o perfil de variabilidade dos componentes voláteis e fixos de 10 diferentes acessos cultivados durante o período de 12 meses. De acordo com os resultados obtidos, o cultivo da espécie foi eficiente, observando-se rápido crescimento, cerca de 1 m de altura a partir do quarto mês de plantio, obtendo-se boa quantidade de biomassa, em média de 400 g por acesso, no período de análise. Com a metodologia desenvolvida para os voláteis foi possível analisar 480 amostras nos 12 meses de amostragem e identificar 14 componentes presentes no óleo das folhas cultivadas. As análises semiquantitativas indicaram que o nerolidol é o volátil majoritário, correspondendo em aproximadamente 50 % do óleo, sendo detectado durante todo ano e, por isso, o mesmo deve ser o maior responsável pelas propriedades odoríferas intensas, as quais são extremamente valorizadas pelas indústrias de fragrâncias e cosméticos. Além disso, com base no perfil de variabilidade dos voláteis, a época de floração foi a menos produtiva em termos de óleo essencial, em média de 0,3 %. Por outro lado, o período de fevereiro a abril foi o mais produtivo, em média de 1 % de óleo a partir das folhas secas. Adicionalmente, dentre os acessos cultivados, somente o 4 não é viável para o cultivo em larga escala. Esta população apresentou diferenças morfológicas em relação às folhas e se destacou pela baixa produtividade de óleo ao longo do ano. A metodologia desenvolvida por CLAE, permitiu distinguir e quantificar simultaneamente 10 padrões cromatográficos (ácido caféico, ácido cumárico, ácido ferúlico, ácido cinâmico, aromadendrina-4?-metil éter, drupanina, isosakuranetina, artepelin C, bacarina e ácido 2,2-dimetilcarboxietenil-2H-1-benzopirânico), os quais podem ser encontrados tanto em amostras hidroetanólicas de alecrim do campo bem como em amostras hidroalcoólicas de própolis verde. Além disso, todas as características de desempenho do método apresentaram resultados aceitáveis, conforme sugeridos pela Anvisa e Inmetro, e podem garantir com segurança e confiabilidade a quantificação dos metabólitos de interesse. Na análise das substâncias fixas, observou-se que o ácido caféico é o componente majoritário, correspondendo em média de 60 % dos fenólicos em análise. O perfil de variabilidade do artepelin C e bacarina foi crescente no decorrer dos meses em estudo, demonstrando teores mais elevados no último trimestre de amostragem, correspondendo em média de 8 e 25 % respectivamente. O flavonóide aromadendrina-4?-metil éter demonstrou concentrações diferenciadas durante o ano. Todavia, este flavonóide juntamente com o ácido caféico podem ser considerados marcadores quimiotaxonômicos da planta cultivada, já que os mesmos foram detectados durante todo ano e em todas as populações. Contudo, considerando a produção, tanto dos voláteis como dos compostos fenólicos, todos os acessos cultivados, com exceção da população 4, podem ser cultivados em larga escala para estes propósitos / Baccharis dracunculifolia is well known for its interaction with insects, mainly Apis mellifera L., for bearing a wide range of secondary metabolites, as well as for displaying many biological activities. Its leaves are punctuated with secretory thricomes rich in secondary metabolites and secretory ducts, which produce and store essential oils. B. dracunculifolia secondary metabolites are used by Apis mellifera to produce ?green? propolis, which is of great importance for pharmaceutical industry, since it displays anticancer, antibacterial, antiinflammatory and antiulcer, among other activities. Considering the displayed biological activities by B. dracunculifolia and the need to standardize the green propolis raw material, the aims of the present work were: a) to develop a gas chromatography analytical methodology to quantify the major volatile constituents in its cultivated populations; b) to develop and validate a HPLC method to quantify the major phenolic compounds present in the aerial parts of B. dracunculifolia, and in the crude green propolis. The developed chromatographic tools were used to study the seasonal influence in the biosynthesis of both volatile oils and phenolic components in 10 different cultivated populations during one year period. All the cultivated populations grew quite well, since it achieved 1 m in height in the fourth month of growth, and it produced an average of 400 g of biomass per individual. Using the developed GC methodology it was possible to analyze 480 samples, and to quantify 14 volatile components of the leaves during one year of its cultivation. The GC analysis indicated that nerolidol was the major compound present in the volatile fraction, about 50 %, which was detected during the entire experiment, and it shall be the most responsible compound for its intense fragrance with importance for cosmetic industry. On one hand, it was observed that during the flowering period the volatile oil production was lower, about 0.3 % w/w, and on the other hand, during the months of February, March and April, its production was higher, about 1 % w/w in dry leaves. Besides, it was observed that population 4 stood out, since it was morphologically different from all others and produced lower yielding of volatile components during the entire year. The HPLC developed methodology allowed to separate and quantify 10 phenolic (caffeic acid, coumaric acid, ferulic acid, cinnamic acid, aromadendrin-4?-methyl ether, drupanin, isosakuranetin, artepillin C, bacharin, 2,2-dimethylcarboxyethenyl-2H-1-benzopyran acid) compounds simultaneously in both B. dracunculifolia and green propolis ethanol extracts. Moreover, all the evaluated validation parameters displayed acceptable results, as established by Brazilian Governmental regulation agencies, Anvisa and Imetro, and it provide very good results for the accurate quantification of phenolic compounds of interest. The analysis indicated that caffeic acid was the major compound present in the analyzed samples, and it corresponded to 60 % of the total quantified phenolic compounds. The contents of artepelin C and bacharin varied greatly during the studied period, displaying higher contents during the last trimester of 8 e 25 %, respectively. The flavonoid aromadendrin-4?-methyl ether has not displayed a coherent pattern during the experiment, since it variable greatly. However, aromadendrin along with caffeic acid could be considered good chemical markers for the analysis of cultivated B. dracunculifolia, considering that both were found in all the samples. Overall, considering the production of both the volatile and phenolic compounds, all the cultivated populations, with exception of the population 4, could be cultivated in large scale for these purposes.

Os extratos de média polaridade de sete indivíduos de Piper solmsianum coletados em diferentes localidades foram analisados por cromatografia líquida de alta eficiência para se verificar diferenças quantitativas na concentração relativa de apiol, dilapiol, miristicina, grandisina, elemicina, isoelemicina e lignana tetraidrofurânica rel- (7R, 8R, 7\'R, 8\'R)-3,4,3\',4\'-dimetilenodioxi-5,5\'-dimetoxi-7,7\'-epoxilignana. Todos os perfis cromatográficos consistiam de apenas dois, três ou quatro picos majoritários. O(s) composto(s) majoritário(s) era(m) distinto(s) de planta para planta, e em um dos espécimes o pico predominante não era nenhum dos sete metabólitos secundários avaliados. Estudaram-se as variações diárias e anuais em três espécimes de uma população selvagem de P. solmsianum localizada na Reserva da CUASO, em São Paulo. A grandisina apresentou teores máximos no início da noite e teores mínimos no início da manhã. Apiol/dilapiol apresentou variação circadianas evidentes apenas em dois indivíduos, sendo que em um deles o teor máximo ocorreu por volta das oito horas da noite, e no outro, o teor máximo ocorreu por volta das cinco horas da manhã. A variação sazonal mostrou pequena amplitude de oscilação. Em dois espécimes o máximo teor de apiol/dilapiol e grandisina ocorreu no verão. No terceiro espécime não se observou variação periódica anual. / The crude extracts of seven individuals of Piper solmsianum collected in different places were analyzed by high efficiency liquid chromatography (HPLC) in order to determine changes in the relative content of apiol, dillapiol, myristicin, grandisin, elemicin, isoelemicin, and tetrahydrofuran lignan rel-(7R,8R,7R,8R)-3,4,3,4- dimethylenedioxy-5,5-dimethoxy-7,7-epoxylignan. All chromatographic profiles consisted of two, three or four major peaks with significant qualitative and quantitative differences. Major components were different from plant to plant and in one specimen an component different from the seven metabolites observed in most cases. The leaf extracts from three wild specimens of P. solmsianum collected at CUASO Reserve, showed annual and diurnal variations in apiol/dillapiol and grandisin relative concentrations. Three plants presented circadian variation with maximum content of grandisin late afternoon, and minimum content in the dawn. Apiol/dillapiol displayed clear circadian variation in two individuals, but without synchrony between phases. The maximum content was observed and in a second specimen around 8 p.m., and in the another specimen, the maximum content occur was observed at 5 a.m. The seasonal variation showed small amplitude of oscillation and in two individuals the maxima contents of apiol/dillapiol and grandisin were observed in summer

Baccharis dracunculifolia é conhecida por sua interação com insetos, especialmente Apis mellifera L., por possuir ampla diversidade de metabólitos secundários, bem como, por apresentar várias atividades biológicas. Suas folhas são pontuadas por tricomas secretores ricos em metabólitos secundários e também apresentam dutos secretores, estruturas que produzem e armazenam os óleos essenciais. Os metabólitos secundários de B. dracunculifolia são usados pelas abelhas A. mellifera para a produção de própolis ?verde?, que é extremamente valorizada pelas indústrias farmacêuticas, pois apresenta atividades antitumoral, antimicrobiana, antiinflamatória, antiulcerogênica, dentre outras. Considerando estas importantes atividades biológicas, e a necessidade de padronização da própolis, os objetivos do presente trabalho foram: desenvolver metodologia analítica por cromatografia gasosa para quantificar os constituintes majoritários presentes no óleo essencial da planta cultivada; desenvolver e validar metodologia analítica por CLAE capaz de garantir a autenticidade dos extratos hidroalcoólicos de B. dracunculifolia, bem como de própolis verde; determinar o perfil de variabilidade dos componentes voláteis e fixos de 10 diferentes acessos cultivados durante o período de 12 meses. De acordo com os resultados obtidos, o cultivo da espécie foi eficiente, observando-se rápido crescimento, cerca de 1 m de altura a partir do quarto mês de plantio, obtendo-se boa quantidade de biomassa, em média de 400 g por acesso, no período de análise. Com a metodologia desenvolvida para os voláteis foi possível analisar 480 amostras nos 12 meses de amostragem e identificar 14 componentes presentes no óleo das folhas cultivadas. As análises semiquantitativas indicaram que o nerolidol é o volátil majoritário, correspondendo em aproximadamente 50 % do óleo, sendo detectado durante todo ano e, por isso, o mesmo deve ser o maior responsável pelas propriedades odoríferas intensas, as quais são extremamente valorizadas pelas indústrias de fragrâncias e cosméticos. Além disso, com base no perfil de variabilidade dos voláteis, a época de floração foi a menos produtiva em termos de óleo essencial, em média de 0,3 %. Por outro lado, o período de fevereiro a abril foi o mais produtivo, em média de 1 % de óleo a partir das folhas secas. Adicionalmente, dentre os acessos cultivados, somente o 4 não é viável para o cultivo em larga escala. Esta população apresentou diferenças morfológicas em relação às folhas e se destacou pela baixa produtividade de óleo ao longo do ano. A metodologia desenvolvida por CLAE, permitiu distinguir e quantificar simultaneamente 10 padrões cromatográficos (ácido caféico, ácido cumárico, ácido ferúlico, ácido cinâmico, aromadendrina-4?-metil éter, drupanina, isosakuranetina, artepelin C, bacarina e ácido 2,2-dimetilcarboxietenil-2H-1-benzopirânico), os quais podem ser encontrados tanto em amostras hidroetanólicas de alecrim do campo bem como em amostras hidroalcoólicas de própolis verde. Além disso, todas as características de desempenho do método apresentaram resultados aceitáveis, conforme sugeridos pela Anvisa e Inmetro, e podem garantir com segurança e confiabilidade a quantificação dos metabólitos de interesse. Na análise das substâncias fixas, observou-se que o ácido caféico é o componente majoritário, correspondendo em média de 60 % dos fenólicos em análise. O perfil de variabilidade do artepelin C e bacarina foi crescente no decorrer dos meses em estudo, demonstrando teores mais elevados no último trimestre de amostragem, correspondendo em média de 8 e 25 % respectivamente. O flavonóide aromadendrina-4?-metil éter demonstrou concentrações diferenciadas durante o ano. Todavia, este flavonóide juntamente com o ácido caféico podem ser considerados marcadores quimiotaxonômicos da planta cultivada, já que os mesmos foram detectados durante todo ano e em todas as populações. Contudo, considerando a produção, tanto dos voláteis como dos compostos fenólicos, todos os acessos cultivados, com exceção da população 4, podem ser cultivados em larga escala para estes propósitos / Baccharis dracunculifolia is well known for its interaction with insects, mainly Apis mellifera L., for bearing a wide range of secondary metabolites, as well as for displaying many biological activities. Its leaves are punctuated with secretory thricomes rich in secondary metabolites and secretory ducts, which produce and store essential oils. B. dracunculifolia secondary metabolites are used by Apis mellifera to produce ?green? propolis, which is of great importance for pharmaceutical industry, since it displays anticancer, antibacterial, antiinflammatory and antiulcer, among other activities. Considering the displayed biological activities by B. dracunculifolia and the need to standardize the green propolis raw material, the aims of the present work were: a) to develop a gas chromatography analytical methodology to quantify the major volatile constituents in its cultivated populations; b) to develop and validate a HPLC method to quantify the major phenolic compounds present in the aerial parts of B. dracunculifolia, and in the crude green propolis. The developed chromatographic tools were used to study the seasonal influence in the biosynthesis of both volatile oils and phenolic components in 10 different cultivated populations during one year period. All the cultivated populations grew quite well, since it achieved 1 m in height in the fourth month of growth, and it produced an average of 400 g of biomass per individual. Using the developed GC methodology it was possible to analyze 480 samples, and to quantify 14 volatile components of the leaves during one year of its cultivation. The GC analysis indicated that nerolidol was the major compound present in the volatile fraction, about 50 %, which was detected during the entire experiment, and it shall be the most responsible compound for its intense fragrance with importance for cosmetic industry. On one hand, it was observed that during the flowering period the volatile oil production was lower, about 0.3 % w/w, and on the other hand, during the months of February, March and April, its production was higher, about 1 % w/w in dry leaves. Besides, it was observed that population 4 stood out, since it was morphologically different from all others and produced lower yielding of volatile components during the entire year. The HPLC developed methodology allowed to separate and quantify 10 phenolic (caffeic acid, coumaric acid, ferulic acid, cinnamic acid, aromadendrin-4?-methyl ether, drupanin, isosakuranetin, artepillin C, bacharin, 2,2-dimethylcarboxyethenyl-2H-1-benzopyran acid) compounds simultaneously in both B. dracunculifolia and green propolis ethanol extracts. Moreover, all the evaluated validation parameters displayed acceptable results, as established by Brazilian Governmental regulation agencies, Anvisa and Imetro, and it provide very good results for the accurate quantification of phenolic compounds of interest. The analysis indicated that caffeic acid was the major compound present in the analyzed samples, and it corresponded to 60 % of the total quantified phenolic compounds. The contents of artepelin C and bacharin varied greatly during the studied period, displaying higher contents during the last trimester of 8 e 25 %, respectively. The flavonoid aromadendrin-4?-methyl ether has not displayed a coherent pattern during the experiment, since it variable greatly. However, aromadendrin along with caffeic acid could be considered good chemical markers for the analysis of cultivated B. dracunculifolia, considering that both were found in all the samples. Overall, considering the production of both the volatile and phenolic compounds, all the cultivated populations, with exception of the population 4, could be cultivated in large scale for these purposes.

Red alder (Alnus rubra) is a tree species with high economic and ecological importance. It is subject to defoliation during unpredictable, episodic outbreaks of tent caterpillars (Malacosoma spp.) that result in reduced growth, decreased wood production, unsightly appearance and mortality in severe cases. Alder trees are weakened by severe and repeated tent caterpillar defoliation, and this can increase the susceptibility of the trees to other pests, diseases and drought. Repeated attack by tent caterpillars can cause decline in red alder populations, which can have potential negative impacts on the ecological and economic benefits of the species. Evidence from other species has shown that plants produce phytochemicals for defense against herbivores at a cost to growth and reproduction, but the relative magnitude of the cost of allocating available resources to defense depends on the level of the resources, and the plant genotype. The quality of a plant as food for herbivores is influenced by leaf physical and biochemical traits, and these traits change during a growing season or upon attack by herbivores. My research aimed to explore the defense mechanisms of red alder against western tent caterpillars (Malacosoma californicum) and determine the resistance variation among and within red alder populations, and to evaluate red alder available resource (nitrogen) allocation to defense and growth. Bioassay feeding trials were conducted in 2014 and 2015 with western tent caterpillars (WTC) (M. californicum) on twenty red alder clones from ten provenances. Phenology and quality of red alder leaves as food for the defoliators were analyzed to determine if budburst, leaf chemical content, water content or physical traits are major determinants of western tent caterpillars preference for red alder leaves. In another experiment, one-year-old seedlings from 100 half-sib red alder families were treated with two levels of ammonium nitrate (NH4NO3) in two growing seasons in a common garden. Growth, herbivore defense-related traits and root nodulation were measured and ranked among the plant genotypes and between the two nitrogen (N) treatments. Leaves from the two N treatments and different alder families were also used for bioassay feeding trials with WTC larvae to determine effects of N and genotype on red alder herbivory resistance. In my final experiments, I harvested and analyzed leaves from three-year-old red alder trees from five different families on eight dates from early April to mid-October 2016 to quantify oregonin and total phenolics concentrations, and wound induction experiments were conducted to determine if the concentrations of the chemicals vary during a growing season and upon attack by insects. Alder clones and families differed in percentage leaf area eaten by caterpillars and in leaf defense traits. The concentrations of foliar phenolic compounds negatively correlated with the percentage leaf area eaten by the caterpillars, but the results suggest a threshold, above which the concentration of each of the chemicals appeared to reduce WTC feeding, individually. Particularly, foliar oregonin concentration above 20 % leaf dry weight consistently appeared to reduce feeding by caterpillars. N availability had significant effects on red alder seedling total dry biomass and leaf N concentration. There was a clear trade-off between red alder seedling growth, and content of the phenolic compounds and leaf thickness, which supports the growth-differentiation balanced hypothesis in relation to resource availability. The concentration of oregonin varied during the growing season and there were no significant responses of any of the measured compounds to wounding. The results suggest that red alder foliar oregonin, condensed tannin and total phenolics are constitutive defenses and are not wound-induced. The effects of leaf total phenolic and condensed tannin concentrations on insect herbivory have been documented by past studies but the effects of oregonin concentration in red alder leaves on tent caterpillar feeding is a novel finding. / Graduate / 2020-04-18

Submitted by (edna.saturno@ufrpe.br) on 2017-02-07T13:28:04Z No. of bitstreams: 1 Ana Paula Oliveira Lucas.pdf: 968479 bytes, checksum: ff02eaef612f94f456a4b1f20ad010f4 (MD5) / Made available in DSpace on 2017-02-07T13:28:04Z (GMT). No. of bitstreams: 1 Ana Paula Oliveira Lucas.pdf: 968479 bytes, checksum: ff02eaef612f94f456a4b1f20ad010f4 (MD5) Previous issue date: 2006-02-17 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / The present work aimed at characterizing the seasonal variation of the zooplankton in the estuaries of the Botafogo and Siriji rivers, northern coast of Pernambuco. The collections were accomplished respectively in May/December 2004, months that included within rainy and dry seasons. Zooplankton samples were collected with net of 300 Qm and 68 Qm mesh size nets, coupled with a flowmeter, to measure the amount of filtered water. In the estuary of the Siriji river, the haulings were accomplished at the surface and above the bottom. Average density logaritmized values in the rainy season varied from 8.24 to 9.35 ind.m-3 in the surface, and 8.71 to 9.47 ind.m-3 at the bottom. In the dry season, these values floated between 7.90 and 9.44 ind.m-3 at the surface, and 4.16 to 9.72 ind.m-3 at the bottom. In the estuary of the Botafogo river, average densities in the rainy season varied from 7.32 to 8.86 ind.m-3 for the macrozooplankton (300 Qm) and, 3.35 to 12.59 ind.m-3 for the microzooplankton (68 Qm). In the dry season, these values floated of 6.14 and 8.40 ind.m-3 for the macrozooplankton and 3.35 to 9.43 ind.m-3 for the microzooplankton. Microzooplankton densities were, most of the time, superior to macrozooplâncton ones, indicating a greater contribution of small individuals (Copepoda nauplii, copepodits and adult). The association analysis showed a total separation of samples collected with different nets (300 and 68 Qm). The most outstanding organisms in the two studied estuaries were Oikopleura dioica, Oikopleura spp., Acartia lilljeborgi, Grapsidae/Ocypodidae/Majidae (zoeae I), Pseudodiaptomus acutus, Oithona oswaldocruzi, Oithona spp. and Copepoda nauplii. Both estuaries presented low species diversity and evenness, thus suggesting signs of environmental disturbances on the structure of the zooplankton community. / O presente trabalho visou caracterizar a variação sazonal do zooplâncton dos estuários dos rios Botafogo e Siriji, litoral norte de Pernambuco. As coletas foram realizadas em maio e novembro/dezembro de 2004, meses que estão incluídos dentro dos períodos chuvoso e seco, respectivamente. As amostras de zooplâncton foram coletadas com rede de 300μm e 68μm de abertura de malha, com fluxômetro acoplado, para medir a quantidade de água filtrada. No estuário do rio Siriji, os arrastos foram realizados na superfície e fundo. Sendo assim, os valores logaritmizados da densidade média na estação chuvosa variaram de 8,24 a 9,35 ind.m-3 na superfície e, 8,71 a 9,47 ind.m-3 no fundo. Na estação seca, estes valores flutuaram entre 7,90 e 9,44 ind.m-3 na superfície e, 4,16 a 9,72 ind.m-3 no fundo. No estuário do rio Botafogo, os valores de densidade média do zooplâncton na estação chuvosa variaram de 7,32 a 8,86 ind.m-3 para o macrozooplâncton (300 μm) e, 3,35 a 12,59 ind.m-3 para o microzooplâcton (68 μm). Na estação seca, estes valores flutuaram de 6,14 e 8,40 ind.m-3 para o macrozooplâncton e 3,35 a 9,43 ind.m-3 para o microzooplâncton. A densidade das amostras de microzooplâncton foi, na maioria das vezes, superior à de macrozooplâncton, indicando uma maior contribuição de pequenos indivíduos(náuplios de Copepoda, copepoditos e espécies adultas). Na análise de associação foi observada a separação total das amostras coletadas com redes distintas (300 e 68 Qm). Os organismos que mais se destacaram nos dois estuários estudados foram Oikopleura dioica, Oikopleura spp., Acartia lilljeborgi, Grapsidae/Ocypodidae/Majidae (zoea I), Pseudodiaptomus acutus, Oithona oswaldocruzi, Oithona spp. e náuplios de Copepoda. Ambos os estuários, apresentaram baixo índice de diversidade e equitabilidade em algumas amostras sugerindo sinais de perturbações ambientais sobre a estrutura da comunidade zooplanctônica.