The Streptomyces are invaluable as a natural source of antibiotics and other bioactive compounds used in medicine and agriculture. S. coelicolor is the model streptomycete, and is studied for its complex secondary metabolism and multicellular life cycle. The subject of this work is bldB, a gene essential for development and antibiotic production in S. coelicolor, and one of its many homologues, located in the abaA antibiotic regulatory locus. The aim was to study the transcriptional regulation of bldB using a luminescent reporter, and investigate the role of each of the genes in the abaA cluster in regulation of antibiotic production, in order to understand the function and mechanism of action of bldB and its homologues. Individual deletion of each of the four genes in the abaA cluster resulted in varying effects on production of the antibiotic CDA. The bldB homologue, SCO0703, was shown to be a positive regulator of CDA, as the null mutant was severely defective in CDA production. It was found that bldB is expressed in most other bld developmental mutants, with the exception of bldD. There was no direct interaction observed between BldD and the bldB promoter, and possible mechanisms of indirect regulation are proposed. / Thesis / Master of Science (MSc)

Submitted in fulfillment of the requirements for the Degree of Master of Technology: Biotechnology, Durban University of Technology, 2009. / Dichrostachys cinerea, Carpobrotus dimidiatus, Capparis tomentosa and Leonotis leonurus are all plants that are indigenous to South Africa. These plants are used in traditional medicine to treat various ailments. However, there is little or no scientific data to justify these traditional uses. Furthermore, it is difficult to reconcile traditional knowledge with scientific evidence because of the overwhelming targeting of signal-responsive systems by plant defensive compounds, multiple sites of action and the connectedness of the signaling pathways, which provide many cures and have pleiotropic effects. In order to evaluate the action spectrum of these plants, and validate its widespread use, this research evaluated the antibacterial, antioxidant, anti-inflammatory, anti-mosquito and immunomodulatory properties of these plants. Antimicrobial activity of the extract was determined by evaluating the bactericidal and fungicidal action using the agar disc diffusion assay. Anti-oxidative properties of the extracts were tested using the DPPH photometric assay. Anti-inflammatory properties were carried out using the 5-lipoxygenase assay. The larvicidal, repellency and insecticidal assay was determined against A.arabiensis. The safe use of these plant extracts was determined by evaluating toxicity, a brine shrimp lethality assay and an in vitro cell culture system using human myelogenous leukemia cell line. Potential carcinogenic activity was evaluated using the Ames Salmonella Mutagenecity assay. The immunomodulatory activity of the extracts on human peripheral blood mononuclear cells 6 was evaluated on freshly harvested lymphocytes using the MTT assay. Cytokine response was evaluated by measuring the secretion of interferon-gamma and interleukin-10. Elucidation of the B cells, T cells, activated T cells, CD 4+, CD 8+ and NK cells was performed by flow cytometry. The extracts showed anti-microbial activity against Escherichia coli, Pseudomonas aeruginosa, Klebsiella oxytoca, Salmonella typhimurium, Serratia marcescens, Bacillus cereus and Tricoderm sp. The highest activity was shown by methanolic and aqueous extracts of L. leonurus leaves followed by methanolic and aqueous extracts of D. cinerea. Extracts of C. tomentosa and D.cinerea demonstrated a higher degree of free radical scavenging than rutin, which was used as a standard indicating that these plants have strong antioxidant properties. None of the plants showed significant anti-inflammatory activity when compared to NDGA. In the anti-mosquito assays, the extracts showed strong repellency and insecticidal activity. L. leonurus extracts demonstrated the highest insecticidal and repellency activity against the mosquito, and was also found to cause ‗knockdown‘ and mortality. The extracts display no toxicity, cytotoxicity and mutagenicity. The immunological studies for immune modulation showed that the methanol extracts of these plants induce a Th1- predominant immune response because they significantly suppressed the secretion of IL-10 and augment IFN-γ production, which are hallmarks used to indicate a stimulation of the innate immune response. This study also provides new information, with respect to the potential use of these plants in producing a mosquito repellent and an immunostimulant. / National Research Foundation

published_or_final_version / abstract / Orthopaedics and Traumatology / Doctoral / Doctor of Philosophy

published_or_final_version / Orthopaedics and Traumatology / Master / Master of Philosophy

No description available.

Orientador: Wagner Vilegas / Resumo: Organismos marinhos são importantes fontes de novos produtos naturais e compostos bioativos com funções variadas. Entre as principais fontes de novas moléculas esta o filo dos Cnidários, com uma importante contribuição no numero de compostos descritos nos últimos anos. Bunodossoma caissarum é uma espécie de anêmona endêmica do Brasil, cujo perfil químico foi analisado. Para essa análise foram realizados uma extração hidroalcoólica e o isolamento de determinadas moléculas com técnicas cromatográficas. Os compostos isolados foram aqueles que apresentaram absorbância no comprimento de onda no UV em 310 nm e foram identificados a partir de técnicas de espectrometria de massas e ressonância magnética. Entre as substâncias encontradas estão sais piridinicos, núcleos imidazóis, caissarona e a etilglicose, moléculas com importantes papeis fisiológicos / Abstract: Marine organisms are importante sources of new natural products and bioactive compounds with various functions. The phylum Cnidaria is among the main sources of new molecules, with na importante contribution to the number of compounds described in the last years. The anemone species Bunodossoma caissarum is endemic to Brazil and had it's chemical profile analyzed. For the purpose of this analysis were done a hydroalcoolic extraction and the isolation of certain molecules by chomatographic techniques. The isolated compounds were those that displayed absorbance in the 310nm wavelength of the UV spectrum and were identified on the basis of mass spectrometry and magnetic ressonance. Among the substances found are pyridinic salts, imidazole nucleus, caissarone and ethyl glucose, which are molecules with important physiological roles / Mestre

by Clifford Sanders. / Thesis (M.S.)--Florida Atlantic University, 2009. / Includes bibliography. / Electronic reproduction. Boca Raton, Fla., 2009. Mode of access: World Wide Web. / Ethylene is the simplest alkene. The carbon-carbon double bond is ubiquitous in the field of chemistry. Ethylene serves as the basis for understanding these molecules. Thus, the assignment of the electronic transitions in ethylene is an important endeavor that many scientists have undertaken, but are yet to decipher theoretically or experimentally. Synchrotron Radiation in the vacuum ultraviolet region allows for magnetic circular dichroism (MCD) measurements of ethylene and other simple alkenes. Studies of ethylene and propylene revealed that the páap* (AgáaB1u ethylene notation) transition is not the lowest energy transition. The páa3s(R) (AgáaB3u ethylene notation) is the lowest energy transition. To further this investigation, MCD and absorption measurement were carried out on isobutene. The isobutene spectra clearly showed four electronic transitions in the 156 to 212 nm wavelength region. These four isobutene transitions have been assigned as páa3s, páap*, páa3p(Sv (Band páa3px proceeding from lower energy to higher energy. The present results support the assignments in ethylene and propylene.

O interesse em buscar fontes naturais para compostos antioxidantes aumentou os estudos sobre frutas, principalmente referente a presença das antocianinas. O camu-camu é um fruto oriundo da região amazônica que vem ganhando destaque nas indústrias de cosméticos, medicamento e alimentos por ser uma fonte rica em antocianinas, ácido ascórbico e minerais. O objetivo do trabalho foi avaliar a incorporação das antocianinas presentes na casca do camu-camu em filmes de desintegração oral à base de gelatina e amido. Os filmes foram produzidos pela técnica casting a partir de blendas de gelatina e amido, com diferentes concentrações (0:100; 25:75; 50:50; 75:25; 100:0), plastificante sorbitol e com a adição de diferentes concentrações do extrato aquoso da casca de camu-camu (0, 10, 15 e 20 g/100g solução filmogênica). Foram analisadas as propriedades mecânicas pelo do teste de tração, cor e opacidade, ângulo de contato, tempo de desintegração, FTIR e MEV. Nos filmes com adição do extrato de camu-camu foram realizadas, adicionalmente, análises de pH de superfície, antocianinas totais e estudo da estabilidade. Com relação as propriedades mecânicas, verificou-se que as alterações nas concentrações de amido e gelatina (AMI:GEL) apresentou diferenças significativas para todos os parâmetros estudados (tensão na ruptura, elongação na ruptura e módulo elástico). O aumento da concentração de gelatina elevou a tensão de ruptura e o aumento da concentração de amido produziu filmes mais frágeis e quebradiços. A adição do extrato apresentou diminuição na tensão de ruptura e no módulo elástico, e aumento na elongação dos filmes. Das análises de cor e opacidade observou-se que com maiores concentrações de amido e do extrato, aumentam os valores da luminosidade, da croma a* e da opacidade, enquanto para a croma b* ocorre uma diminuição. Os ângulos de contato dos filmes diminuíram significativamente com o aumento da concentração de amido e aumentaram com maiores concentrações de extrato. Os filmes com extrato, apresentaram tempo de desintegração inferior, sendo observados tempos maiores com o aumento da concentração de gelatina. Com relação a estabilidade, os filmes apresentaram elevada perda das quantidades de antocianinas para todas as formulações analisadas. Todos os filmes analisados apresentaram incorporação eficiente do extrato, dessa forma, conclui-se que a produção de filmes de desintegração oral com extrato de camu-camu é uma alternativa eficiente para o aproveitamento dos compostos funcionais presentes da casca. / The interest in finding natural sources for antioxidants increased studies on fruit, especially regarding the presence of anthocyanins. Camu-camu is a fruit originating in the Amazon region that is gaining prominence in the cosmetics, medicine and food to be a rich source of anthocyanins, ascorbic acid and minerals. The objective was to evaluate the incorporation of anthocyanins present in the shell of the camu-camu in films orally disintegrating based gelatin and starch. The films were produced by casting from blends of gelatin and starch with different concentrations (0: 100, 25:75, 50:50, 75:25, 100: 0), plasticizer sorbitol and with the addition of different concentrations the aqueous extract of camu-camu peel (0, 10,15 and 20 g / 100 g film solution). The mechanical properties were analyzed using the tensile test, color, and opacity, contact angle, disintegration time, FTIR and SEM. In the films with the addition of camu-camu extract were performed, additionally, surface pH analysis, anthocyanins and stability. Regarding the mechanical properties, it has been found that changes in starch and gelatin concentrations (AMI:GEL) showed significant differences for all the studied parameters (stress, elongation and elastic modulus). The increase in the gelatin concentration to increased breakdown voltage and increased concentration of starch produced more fragile and brittle films. The addition of the extract showed a decrease in breakdown voltage and elastic modulus, and increased elongation of the films. Analysis of the color and opacity was observed that with higher concentrations of starch and the extract, increases the brightness values of a* chroma and opacity as to chroma b* is decreased. The contact angles of the films decreased significantly with increasing starch concentration and increased with higher extract concentrations. The films to extract showed disintegration time lower, longer times being observed with increasing concentration of gelatin. Regarding stability, the films showed high loss of the amounts of anthocyanins for all formulations analyzed. All samples show efficient incorporation films extract thus concluded that the production of the orally disintegrable films with camu-camu extract is an effective alternative to the use of compounds of the functional shell.

As perspectivas são promissoras para a exploração de espécies não tradicionais de frutas tropicais com compostos de interesse nutricional e capacidade antioxidante. Dentre as frutas que apresentam relevantes quantidades de nutrientes, podemos destacar o pequi (Caryocar villosum Alb. Pers.). Na sua forma in natura, possui concentrações expressivas de compostos fenólicos e carotenoides totais, principalmente na polpa, sendo o óleo extraído rico em ácidos graxos. Diversos estudos sobre o cacau relacionam sua alta capacidade antioxidante com sua grande concentração de compostos fenólicos. O objetivo deste estudo foi a utilização do mesocarpo do pequi liofilizado na incorporação em formulação de chocolate amargo com vista à agregação de valor nutricional. No mesocarpo do pequi liofilizado foi verificada a granulometria, biometria, composição centesimal, carotenoides e microscopia eletrônica de varredura. No óleo extraído do mesocarpo do pequi liofilizado foi avaliada a composição em ácidos graxos e colorimetria, bem como a qualidade e funcionalidade nos perfis lipídicos pelos índices de composição: índice de Aterogenicidade (I.A), índice de Trombogenicidade (I.T) e a razão hipocolesterolêmica e hipercolesterolêmica (H.H). Adicionou-se 1,5% de mesocarpo liofilizado na formulação de chocolate amargo determinando-se tamanho de partícula, índice de temperagem, parâmetros físico-químicos, reologia, análise térmica, capacidade antioxidante total pelos métodos de redução do ferro e DPPH, e determinação de fenólicos totais. Como resultado observou-se principalmente a presença dos ácidos graxos palmítíco (26,5 %), oleico (52,67 %) e linoleico (15,2 %) no óleo da polpa. Com a adição de 1,5 % de mesocarpo de pequi liofilizado não houve aumento significativo (p<0,05) do tamanho de partículas da massa do chocolate formulado, não aumentando sua viscosidade e a tensão inicial. A análise de fenólicos totais mostrou um aumento da concentração destes compostos na formulação de chocolate com adição do mesocarpo do pequi liofilizado (813,49 &#181;g GAE.mL -1) em relação à formulação padrão de chocolate amargo (235,98 &#181;g GAE.mL -1), mostrando o grande potencial que este fruto oferece. Na análise de atividade antioxidante, frente ao radical DPPH, houve um aumento da concentração de antioxidantes na formulação de chocolate com adição do mesocarpo do pequi liofilizado (151,65 mg/L) em relação à formulação padrão de chocolate amargo (158,87 mg/L). Na curva de resfriamento da manteiga de cacau encontrou-se o valor BCI (Buhler Crystallization Index) de 3,7 &#177; 0,00, sendo considerado adequado para produção de chocolates. Houve um aumento da temperatura inicial da temperagem e uma diminuição do tempo total do processo com a adição do óleo de pequi na massa do chocolate em relação à formulação controle. A análise térmica indicou que o óleo de pequi suporta temperaturas de até 300 &#176;C, e os chocolates até 50&#176;C. Os índices de funcionalidade do óleo de pequi demostraram que o mesmo possui baixa capacidade de aterogenicidade (0,38), a relação poliinsaturado/ saturado apresentou-se na faixa indicada como benéfica à saúde (0,61), assim como o índice hipocolesterolêmico/ hipercolesterolêmico que se apresentou adequado (2,58). Concluindo-se que houve o aumento no potencial nutricional do chocolate com a adição da polpa de pequi liofilizada, sem alterar estatisticamente suas propriedades reológicas. / The perspectives are promisíng for the exploration of non-traditional species of tropical fruits with compounds of nutritional interest and antioxidant capacity. Among the fruits that present relevant amounts of nutrients, we can highlight pequi (Caryocar villosum Alb. Pers.). In its natural form, it has expressive concentrations of phenolic compounds and total carotenoids, mainly in the pulp, being the extracted oil rich in fatty acids. Several studies about cocoa relate its high antioxidant capacity to its high concentration of phenolic compounds. The objective of this study was the utilization of freeze-dried mesocarp of pequi in the incorporation on bitter chocolate formulation with a view to aggregating nutritional value. In the freeze-dried mesocarp of pequi was verified the granulometry, biometry, centesimal composition, carotenoids and scanning electron microscopy. In the extracted oil of freeze-dried mesocarp of pequi was evaluated the composition of fatty acids and colorimetry, as well the quality and the functionality in the lipids profile by the index of composition: Aterogenicity Index (A.I.), Thrombogenicity Index (T.I.) and hypocholesterolemic and hypercholesterolemic ratio (H.H.). It was added 1.5% of freeze-dried mesocarp in the formulation of bitter chocolate determining particle size, temper index, physico-chemical parameters, rheology, thermal analysis, total antioxidants capacity by reduction of iron and DPPH methods, and determination of total phenolics. As a result, it was observed the main presence of palmitic (26.5%), oleic (52.67%) and linoleic (15.2%) acids in the pulp of the oil. With the addition of 1.5 % of freeze-dried mesocarp of pequi there was no significant increase (p<0.05) in the particle size of the formulated chocolate mass, without increasing its viscosity and initial tension. The analysis of total phenolics showed an increase in the concentration of these compounds in the chocolate formulation with the addition of the freeze-dried mesocarp (813.49 &#181;g GAE.mL-1) in relation to the standard bitter chocolate formulation (235.98 &#181;g GAE.mL-1), showing the great potential that this fruit offers. In the analysis of antioxidant activity, against the radical DPPH, there was an increase in the concentration of antioxidants in the chocolate formulation with the addition of the freeze-dried mesocarp (151,65 mg/L) in relation to the standard bitter chocolate formulation (158.87 mg/L). In the cooling curve of cocoa butter it was found the BCI value (Buhler Crystallization Index) of 3.7 &#177; 0.00 being considered suitable for manufacturing of chocolates. It was occurred an increase of the initial temperature of tempering and a decrease of the total processes time with the addition of pequi Gil in the chocolate mass. The thermal analysis indicated that pequi Gil supports temperatures of up to 300&#176;C, and the chocolates up to 50&#176;C. The functional indexes of pequi Gil showed that it had a low atherogenic capacity (0.38), the polyunsaturated / saturated ratio was in the range indicated as beneficial to health (0.61), as well as the index hypocholesterolemic/ Hypercholesterolemic was adequate (2,58). It was concluded that there was an increase in the nutritional potential of the chocolate with the addition of lyophilized pequi pulp, without statistically altering its rheological properties.

A nutrição possui papel de fundamental importância na promoção da saúde. Os alimentos in natura podem ser encontrados na forma de frutas e vegetais minimamente processados (fresh-cut) embalados. Uma alternativa para a administração de alguns alimentos in natura aos indivíduos imunocomprometidos e neutropênicos, ou até mesmo para a população em geral, é a aplicação de radiação gama, em doses sub-esterilizantes, com o intuito de oferecer alimentos de qualidade e seguros do ponto de vista microbiológico. A descrição na literatura sobre tentativas de oferecer alimentos seguros e saudáveis aos indivíduos portadores de patologias do sistema imune é escassa. Assim, o objetivo do trabalho foi avaliar o efeito de diferentes doses de radiação gama na composição química, no teor de compostos bioativos e atividade antioxidante de abacaxi, salada de frutas, mini cenouras e salada de folhas minimamente processados destinados a dietas especiais. As amostras foram submetidas à radiação gama, com doses de 0.5, 1, 2 e 3 kGy e analisadas juntamente com as amostras controle (não irradiadas). Foram analisados a composição centesimal, compostos fenólicos, flavonóides, proantocianidinas, atividade antioxidante por ORAC e DPPH&bull;, vitamina C e carotenóides. Não foram observadas alterações significativas nos compostos avaliados antes e após a irradiação, com exceção dos carotenóides que demonstraram um aumento significativo dos teores após a aplicação de radiação gama. Assim, pode-se concluir que a irradiação, aplicada em baixas doses e em alimentos minimamente processados, promove a manutenção do valor nutricional do alimento, trazendo benefícios aos indivíduos que necessitam de alimentos seguros e de suficiente aporte nutricional que garanta a promoção e recuperação da saúde, tais como os imunocomprometidos. / Nutrition plays a fundamental role in promoting health. The fresh food can be found in form of minimally processed fruits and vegetables (fresh-cut). An alternative of fresh food administration to immunocompromised and neutropenic individuals, or even to the general population, is the application of gamma radiation in sub-sterilizing doses, in order to provide quality and safe food in microbiological point of view. The description in the literature on attempts to provide safe and healthy food to individuals with immune system disorders is scarce. The objective of the study was to evaluate the effect of different doses of gamma radiation in the chemical composition, the bioactive compounds content and antioxidant activity of minimally processed pineapple, fruit salad, mini carrots and leaves salad for special diets. Samples were subjected to gamma radiation doses of 0.5, 1, 2 to 3 kGy and analyzed at the same time with control samples (not irradiated). There were analyzed the chemical composition, phenolic compounds, flavonoids, proanthocyanidins, antioxidant activity by ORAC and DPPH&bull;, vitamin C and carotenoids. No significant changes were observed in compounds contents before and after irradiation, except carotenoids that have demonstrated a significant increase in levels after application of gamma radiation. Thus, it can be concluded that irradiation applied in low doses and in minimally processed foods promotes the maintenance of food nutritional value, bringing benefits to the individuals who need safe food and adequate nutritional supply to ensure the promotion and recovery health, such as immunocompromised.