Cyanobacterial secondary metabolites are a rich source of bioactive compounds with potential utilization in pharmacology. The aim of this study was to evaluate the effect of the novel compound nostatin A isolated from the cyanobacterium Desmonostoc muscorum. Project was focused on the extraction, purification and characterization of the cytostatic effect caused by this novel compound and its naturally occurred structural analogues. The cytostatic activity of nostatin A and its analogs was evaluated in HeLa cell line. Experiments based on microscopy, flow cytometry and HPLC-HRMS techniques were performed in order to clarify the cytostatic effect of nostatin A in HeLa cells and its mechanism of the action.

Rhodococci are notable for their ability to degrade a variety of natural and xenobiotic compounds. Recently, interest in Rhodococcus has increased due to the discovery of a large number of genes for secondary metabolism. Only a few secondary metabolites have been characterized from the rhodococci (including 3 recently described antibiotics). Twenty-four new Rhodococcus strains were isolated from soils in East Tennessee using acetonitrile enrichment culturing and identified using 16S rRNA analysis. Forty-seven Rhodococcus strains were screened for antibiotic production using a growth inhibition assay. One strain, MTM3W5.2, had 90% similarity to the Rhodococcus opacus 16S rRNA gene sequence and produced a large zone of inhibition against R. erythropolis and a large number of closely related species. The antimicrobial compound produced by MTM3W5.2 had a large MW of 911.5452 Da and acts much like a bacteriocin but no amino acids were detected in this molecule based on TLC analysis.

Peppers are a rich source of diverse bioactive compounds with potential health-promoting properties. The levels of bioactive compounds and antioxidant activity can be affected by analytical methods, pre-harvest factors, and the quality of peppers. In order to understand the nutrient composition and antioxidant activity in peppers, determination of factors influencing the contents of bioactive compounds is important. The overall objectives were to determine the efficient conditions of sample preparation and the impact of pre-harvest factors affecting bioactive compounds and antioxidant activity. Optimal extraction procedures were developed, and HPLC methods were validated for bioactive compounds in peppers. The highest flavonoids were extracted in ethanol, while myricetin was extracted using N-N-dimethylformamide. Optimized conditions for flavonoids were obtained during 3 h of extraction time and hydrolysis in M HCl for 60 min at 95 degrees C. Capsaicinoids and ascorbic acid were simultaneously separated and extracted using a solvent mixture consisting of 3% metaphosphoric acid: ethanol (2:8) after 30 min of sonication. To determine the relationship between bioactive compounds and antioxidant activities in pepper extracts from different solvent properties, bioactive compounds were analyzed, and the antioxidant activities were assayed by 2,2,-Diphenyl-1-picryl hydrazyl (DPPH), reducing power, and degradation of deoxyribose. Hexane extracts had the highest levels of capsaicinoids and carotenoids, while methanol extracts had the highest levels of flavonoids. Strong DPPH scavenging activity and reducing power were found in lipophilic extracts, while hydrophilic extracts were appropriate for inhibition of deoxyribose degradation. Variation in content of ascorbic acid, capsaicinoids, and flavonoids was evaluated at immature and mature stages of pepper cultivars in different locations over two years. Mature peppers contained the highest levels of capsaicinoids and ascorbic acid. Flavonoids were variable at different maturity stages. Interactions between pre-harvest factors and bioactive compounds were highly significant. This study demonstrated the efficient sample preparation methods and simultaneous separation of bioactive compounds, which reduces analysis time and leads to reduced cost. The antioxidant properties were strongly associated with the concentration of bioactive compounds based on selective pepper extracts. The pepper quality can be improved by using appropriate pre-harvest conditions that increase the levels of bioactive compounds in peppers.

Colorectal cancer is the third most common cancer and the fourth most common cause of cancer-related death in the world. The global burden of colorectal cancer is also expected to increase by 60%, to over 2.2 million new cases and 1.1 million annual deaths, by the year 2030. Jackfruit is known for its packed nutrition including many antioxidants: vitamin C, carotenoids and flavanones. It has also been used in traditional medicine due to its potential protection against many chronic diseases. However, there is limited research studying the potential effect of jackfruit on colorectal cancer. Here, we used a well-established AOM/DSS mice model to investigate the impact of jackfruit-derived extracts on colitis-associated colorectal cancer. After 6-week treatment with diet containing 480 ppm jackfruit-derived extracts, the mice showed significantly alleviated colon tumorigenesis with a 46% decrease in tumor numbers of each mouse compared to vehicle group (2.1 ± 0.31 for 480 ppm jackfruit-derived fraction group vs 3.9 ± 0.67 for vehicle group, P < 0.05). The expression of the pro-inflammatory cytokines (Il-6 and Inf- γ) and pro-tumorigenic genes (Axin2, Vegf, Myc and Pcna) was also decreased in the group consuming 480 ppm jackfruit-derived extracts compared to the vehicle group. Together the results suggest that the consumption of jackfruit-derived extracts could protect against colitis-associated colorectal carcinogenesis in mice.

Orientador: Ana Paula Girol / Resumo: A poluição do ar e o uso de cigarro são as principais fontes de inalação e exposição a agentes carcinógenos, como os hidrocarbonetos aromáticos policíclicos (HAP) resultantes da combustão incompleta de matéria orgânica e distribuídos amplamente no ambiente. O benzopireno é um dos principais HAP presentes no ambiente e com alta capacidade carcinogênica, sendo, portanto, usado em modelos experimentais in vivo. Investigações têm mostrado que os fármacos anti-inflamatórios podem reduzir a incidência de câncer de pulmão. Os compostos bioativos naturais (CBN) como os terpenos e flavonoides apresentam muitas propriedades, entre elas, redução da inflamação, estimulação do sistema imune e modulação de enzimas detoxificantes. Entre os CBNs destacamos a piperlongumina (PL), um alcaloide da Piper longum, que pode representar uma possibilidade de tratamento para o câncer de pulmão. Para o desenvolvimento do trabalho foram usados camundongos Balb/c divididos em 3 grupos (n=10/grupo): controle (sham), induzido por benzopireno (100 mg/kg, diluído em DMSO e administrado intraperitonealmente, i.p.) não tratado (BaP) e induzido por benzopireno e tratado com PL (2 mg/kg em DMSO a 10%, i.p.) a partir da oitava semana pós-indução (BaP/PL). Os animais foram pesados diariamente. A pletsmografia para verificação de volume, frequência e ventilação pulmonares foi realizada na última semana do protocolo (12ª semana). Após, os animais foram eutanasiados para coleta de sangue, lavado broncoalveolar e frag... (Resumo completo, clicar acesso eletrônico abaixo) / Mestre

The soil bacterium Rhodococcus has a wide array of secondary metabolic pathways such as production of pigments, siderophores, and antibiotics that makes it an organism of interest for the production of novel natural products. Analysis of the genome sequence of Rhodococcus indicates the presence of 24 non-ribosomal peptide synthases and 7 polyketide synthases possibly involved in production of secondary metabolites. The use of a solid agar extraction method to screen soil isolates of Rhodococcus for compounds with inhibitory activity against other bacteria resulted in the discovery of a promising candidate molecule. The Rhodococcus strain KCHXC3, isolated from eastern Tennessee soil, produces a substance that inhibits the growth of several Gram-negative bacteria such as Escherichia coli, Pseudomonas aeruginosa, and Klebsiella pneumonia and Gram-positive bacteria such as Micrococcus luteus and, Staphylococcus aureus. After bulk extraction of this compound with ethyl acetate from agar plates, the material was partially purified through different chromatography processes.

Streptomycetes are soil filamentous Gram-positive bacteria that produce wide variety of pigments and biologically active substances including macrolides. Some of them are used as very efficient antibiotics and strong antifungal agents in medicine, others have became useful tools for staining biomembranes and detecting cholesterol via their internal fluorescence. Actinomycete Streptomyces durmitorensis (wild type strain MS405T ) is a bacteria isolated from Durmitor National Park in Montenegro soil samples. It produces secondary metabolite that has been identified as 32,33-didehydroroflamycoin (DDHR) closely related to the macrolides roflamycoin and generaly used filipin. DDHR exhibits cytototoxic activity against mammalian cells and yeast Saccharomyces cerevisiae strain EGY48. In addition it has interesting fluorescence properties allowing visualization of some membrane components. DDHR interacts with biomembranes, causes their disintegration leading to changes of the actin and tubulin cytoskeleton organization and in higher concentrations it causes cells necrosis. DDHR-sterol interaction in cell membranes decreases fluorescence intensity of DDHR. The compound is able to fluorescently stain aberrant lysosomes and could be therefore potentially used in diagnostics of some lysosomal storage disease.

Submitted by (edna.saturno@ufrpe.br) on 2016-07-26T11:49:42Z No. of bitstreams: 1 Ana Carla da Silva Caetano.pdf: 782522 bytes, checksum: f2c222ad97acdbff5efc95970a75c528 (MD5) / Made available in DSpace on 2016-07-26T11:49:42Z (GMT). No. of bitstreams: 1 Ana Carla da Silva Caetano.pdf: 782522 bytes, checksum: f2c222ad97acdbff5efc95970a75c528 (MD5) Previous issue date: 2009-02-26 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The observation that the plants have biologically active substances, mainly polyphenols, that provide benefits to health, has driven studies on the antioxidant property. Thus, considering that the agri-industrial waste from acerola also contains phenolic compounds, this study was carried out to determine the procedure for extraction of polyphenols and the antioxidant activity of their extracts. Hyidroacetone, hydroethanolic, hydromethanolic and aqueous extracts, obtained by sequential extraction procedure, combining three extraction cycles (20 minutes each one) and two temperatures (25 ± 2ºC and 50 ± 2ºC), were submitted to the determination of total phenolic and screened for their free DPPH• (1,1-diphenyl-2-picrilhidrazil) and ABTS•+ (2,2 '-azino-bis-(3-etilbenzotiazolina 6-sulfonic acid) scavenging activity and their effect on the linoleic acid peroxidation by the ferric thiocyanate method.Soybean oil with the addition of the extracts (200ppm) were submitted to anaccelerated storage test in an oven at 60°C for 28 days, where sample were taken at time intervals and analyzed for peroxide value and conjugated dienes. The extracts showed high content of total polyphenols; however, the hydroacetone extract exhibited the highest total phenolic content (5,954.2 μg catechin equivalent. mL-1). Hydroethanolic and hydromethanolic extracts exhibited good DPPH• scavenging activity (low value of EC50 and TEC50, high value of EA), good ABTS•+ scavenging capacity (1.445,1 e 1.145,5μMol TEAC/g, respectively), high percentage of inhibition of peroxidation of linoleic acid (96.12% and 91.84%, respectively). These extracts showed the ability to retard the formation of peroxides and conjugated dienes in soybean oil. The procedure involves aqueous-organic solvents, three cycles of extraction and temperature of 25°C is effective in the extraction of phenolic compounds. Although, the hydroacetone extract has exhibited the highest total phenolic content, the antioxidant action of this extract was less than shown by hydroethanolic and hydromethanolic extracts that, under the experimental conditions, exhibited the best antioxidant activity, both in model system and in soybean oil. Therefore, the agri-industrial waste from acerola can be considered as a source of natural antioxidants and can be useful in food applications in replace or in association with synthetic antioxidants. / A constatação de que os vegetais possuem substâncias biologicamente ativas, principalmente polifenóis, que trazem benefícios à saúde, tem impulsionado estudos sobre sua propriedade antioxidante. Assim, considerando que os resíduos gerados pela agroindústria de polpa de frutas, ainda, contém significante teores de compostos fenólicos, este trabalho teve como objetivo determinar o solvente e a temperatura mais eficiente para a extração destes fitoquímicos a partir de resíduos de acerola e avaliar o potencial antioxidante de seus extratos. Desta forma, extratos hidroacetônico,hidroetanólico, hidrometanólico e aquoso, obtidos por processo de extração seqüencial, em duas temperaturas (25 ± 2ºC e 50 ± 2ºC), foram avaliados quanto ao teor de compostos fenólicos, e a sua ação antioxidante em sistemas modelos, por meio do método da capacidade de seqüestro do radical DPPH• (1,1-difenil-2-picrilhidrazil), do radical ABTS•+ (2,2’-azino-bis- (3-etilbenzotiazolina-6-ácido sulfônico) e da inibição da peroxidação do ácido linoléico pelo método Tiocianato Férrico, bem como, em óleo de soja por meio do teste acelerado em estufa. Quanto ao teor de fenólicos, independente da temperatura usada no processo de extração, destacou-se o hidroacetônico por ter apresentado o mais elevado teor (5.954,2 μg em equivalente de catequina. mL-1). Os extratos hidroetanólicos e hidrometanólico exibiram forte capacidade de seqüestro do radical DPPH• (baixo valor de EC50 e de TEC50, alto valor de EA), elevada capacidade de seqüestro do radical ABTS•+ (1.445,1 e 1.145,5μMol TEAC/g, respectivamente), e forte proteção do ácido linoléico contra a peroxidação (96,12% e 91,84% respectivamente). Estes extratos, também, apresentaram boa capacidade em retardar formação de peróxidos e de dienos conjugados em óleo de soja. Os resultados obtidos permitem evidenciar que o procedimento envolvendo solventes orgânico-aquosos, três ciclos de extração e a temperatura de 25°C se mostrou eficaz na extração de fenólicos totais. O solvente hidroacetônico embora tenha propiciadoa extração de uma maior quantidade de fenólicos totais, a ação antioxidante deste extrato foi inferior a exibida pelos extratos hidroetanólico e hidrometanólico que, nas condições experimentais, exibiram a melhor ação antioxidante, tanto em sistema modelo como no teste acelerado em estufa. Desta forma, o resíduo agroindustrial de acerola apresenta-se como fonte de antioxidante natural, permitindo vislumbrar a sua utilização em alimentos em substituição ou em associação aos antioxidantes sintéticos.

A biodiversidade brasileira é conhecida devido a sua riqueza de espécies, sendo considerada uma fonte promissora de produtos naturais. Dentre as plantas vasculares, a família Solanaceae A. Juss. (Solanaceae) é considerada uma das maiores, apresentando distribuição em todas as regiões tropicais e temperadas do mundo. A família Solanaceae apresenta alta diversidade de espécies de importância econômica como fonte de alimentos, propriedades medicinais e ornamentais. Plantas desta família são fontes de metabólitos secundários de diversas classes químicas com as mais diversas aplicações. Fungos fitopatogênicos são responsáveis por causar diversas doenças e consideráveis perdas na agricultura. O controle das doenças é realizado através de métodos químicos, físicos e biológicos, porém o uso excessivo e ininterrupto de produtos químicos pode resultar na seleção de micro-organismos resistentes. Além disto, muitos fungicidas apresentam toxicidade alta e a sua utilização indiscriminada pode causar efeitos indesejáveis sobre outros organismos no ambiente. O objetivo deste estudo é explorar o potencial biológico e químico de metabólitos secundários produzidos por plantas da família Solanaceae com potencial fungitóxico a fitopatógenos. Foram selecionadas 15 espécies de plantas da família Solanaceae que tiveram os extratos de suas folhas avaliados em ensaios biológicos in vitro sobre o crescimento micelial de 6 fitopatógenos de importância na agricultura. Dentre estas, três foram selecionadas para uma investigação mais aprofundada, as espécies Solanum americanum, Acnistus arborescens e Physalis peruviana. No estudo da planta S. americanum, foram identificados compostos bioativos pertencentes à classe dos glicoalcalóides, que inibiram o crescimento micelial do fitopatógeno Moniliophthora perniciosa. No estudo de A. arborescens foi identificado a presença de um composto ativo pertencente à classe dos vitanolidos, provavelmente o 7&beta;-acetoxivitanolido D, e sua ação antifúngica está sendo relatada pela primeira vez. No estudo da planta P. peruviana a fração semipura bioativa indicou a presença de um composto pertencente à classe dos vitanolidos. Esses resultados evidenciaram que os compostos presentes nas plantas, apresentam bioatividade que inibem o crescimento micelial dos fungos fitopatogênicos Moniliophthora perniciosa e Phytophthora cinnamomi, podendo ser uma nova opção coadjuvante no controle de fitopatógenos. / The Brazilian biodiversity is known due to its richness of species, and is considered a promising source of natural products. Among the vascular plants, the family Solanaceae A. Juss. (Solanaceae) is considered one of the largest, with distribution in all tropical and temperate regions of the world. The Solanaceae family has a high diversity of species of economic importance as a source of food, medicinal and ornamental properties. Plants of this family are sources of secondary metabolites from different chemical classes with many different applications. Plant fungi are responsible for causing various diseases and considerable losses in agriculture. The disease control is accomplished through chemical, physical and biological methods, but the excessive and continuous use of chemical products, may result in selection of resistant micro-organisms, in addition, many fungicides have a high toxicity and its indiscriminate use can cause undesirable effects on other organisms in the environment. The objective of this study is to explore the biological and chemical potential of secondary metabolites produced by plants of the Solanaceae family with potential fungitoxic the pathogens. We selected 15 species of Solanaceae plants that had the extracts of its leaves evaluated in vitro biological assays on the mycelial growth of 6 plant pathogens of importance in agriculture. Among these, three were selected for further investigation, the species Solanum americanum, Physalis peruviana and Acnistus arborescens. In the study of plant S. americanum, it was identified bioactive compounds belonging to the class of glycoalkaloids that inhibited the mycelial growth of the pathogen M. perniciosa. In the study of A. arborescens it was identified the presence of the active compound belonging to the class of withanolides, probably the 7&beta;-acetoxywithanolide D, and its antifungal activity is being reported for the first time. In the study of plant P. peruviana semipure bioactive fraction indicated the presence of the compound belonging to the class of withanolides. These results showed that the compounds present in plants, have bioactivity that inhibit the mycelial growth of pathogenic fungi M. perniciosa and P. cinnamomi and may be a new option in the adjuvant control pathogens.

A biodiversidade brasileira é conhecida devido a sua riqueza de espécies, sendo considerada uma fonte promissora de produtos naturais. Dentre as plantas vasculares, a família Solanaceae A. Juss. (Solanaceae) é considerada uma das maiores, apresentando distribuição em todas as regiões tropicais e temperadas do mundo. A família Solanaceae apresenta alta diversidade de espécies de importância econômica como fonte de alimentos, propriedades medicinais e ornamentais. Plantas desta família são fontes de metabólitos secundários de diversas classes químicas com as mais diversas aplicações. Fungos fitopatogênicos são responsáveis por causar diversas doenças e consideráveis perdas na agricultura. O controle das doenças é realizado através de métodos químicos, físicos e biológicos, porém o uso excessivo e ininterrupto de produtos químicos pode resultar na seleção de micro-organismos resistentes. Além disto, muitos fungicidas apresentam toxicidade alta e a sua utilização indiscriminada pode causar efeitos indesejáveis sobre outros organismos no ambiente. O objetivo deste estudo é explorar o potencial biológico e químico de metabólitos secundários produzidos por plantas da família Solanaceae com potencial fungitóxico a fitopatógenos. Foram selecionadas 15 espécies de plantas da família Solanaceae que tiveram os extratos de suas folhas avaliados em ensaios biológicos in vitro sobre o crescimento micelial de 6 fitopatógenos de importância na agricultura. Dentre estas, três foram selecionadas para uma investigação mais aprofundada, as espécies Solanum americanum, Acnistus arborescens e Physalis peruviana. No estudo da planta S. americanum, foram identificados compostos bioativos pertencentes à classe dos glicoalcalóides, que inibiram o crescimento micelial do fitopatógeno Moniliophthora perniciosa. No estudo de A. arborescens foi identificado a presença de um composto ativo pertencente à classe dos vitanolidos, provavelmente o 7&beta;-acetoxivitanolido D, e sua ação antifúngica está sendo relatada pela primeira vez. No estudo da planta P. peruviana a fração semipura bioativa indicou a presença de um composto pertencente à classe dos vitanolidos. Esses resultados evidenciaram que os compostos presentes nas plantas, apresentam bioatividade que inibem o crescimento micelial dos fungos fitopatogênicos Moniliophthora perniciosa e Phytophthora cinnamomi, podendo ser uma nova opção coadjuvante no controle de fitopatógenos. / The Brazilian biodiversity is known due to its richness of species, and is considered a promising source of natural products. Among the vascular plants, the family Solanaceae A. Juss. (Solanaceae) is considered one of the largest, with distribution in all tropical and temperate regions of the world. The Solanaceae family has a high diversity of species of economic importance as a source of food, medicinal and ornamental properties. Plants of this family are sources of secondary metabolites from different chemical classes with many different applications. Plant fungi are responsible for causing various diseases and considerable losses in agriculture. The disease control is accomplished through chemical, physical and biological methods, but the excessive and continuous use of chemical products, may result in selection of resistant micro-organisms, in addition, many fungicides have a high toxicity and its indiscriminate use can cause undesirable effects on other organisms in the environment. The objective of this study is to explore the biological and chemical potential of secondary metabolites produced by plants of the Solanaceae family with potential fungitoxic the pathogens. We selected 15 species of Solanaceae plants that had the extracts of its leaves evaluated in vitro biological assays on the mycelial growth of 6 plant pathogens of importance in agriculture. Among these, three were selected for further investigation, the species Solanum americanum, Physalis peruviana and Acnistus arborescens. In the study of plant S. americanum, it was identified bioactive compounds belonging to the class of glycoalkaloids that inhibited the mycelial growth of the pathogen M. perniciosa. In the study of A. arborescens it was identified the presence of the active compound belonging to the class of withanolides, probably the 7&beta;-acetoxywithanolide D, and its antifungal activity is being reported for the first time. In the study of plant P. peruviana semipure bioactive fraction indicated the presence of the compound belonging to the class of withanolides. These results showed that the compounds present in plants, have bioactivity that inhibit the mycelial growth of pathogenic fungi M. perniciosa and P. cinnamomi and may be a new option in the adjuvant control pathogens.