Venereal diseases (VDs) are infections that are mainly transmitted through sexual intercourse and amongst these are gonorrhoea, syphilis, chlamydia and trichomoniasis. Gonorrhoea is the most commonly known VD and the widest spread contagious infection in the world. Out of 448 million cases of curable venereal infections, gonorrhoea represents 88 million cases and the rest are syphilis, chlamydia and trichomoniasis. Gonorrhoea has recently been rated as in the emergent multidrug resistance phase. Venereal diseases are amongst the major diseases ravaging many rural communities. People infected with these diseases are considered a disgrace in the community. Indigenous populations, for example the Vha-Venda people tend to use medicinal plants to treat these infectious diseases rather than using western medicines. Vha-Venda people have depended on medicinal plants for their health and survival for millenia. In order to validate and give scientific credence to the use of medicinal plants by the Vha-Venda people for venereal diseases, several pharmacological assays were carried out. The study was aimed at evaluating the; antimicrobial, anti-inflammatory activities, HIV-type 1 reverse transcriptase (RT) inhibition properties and to determine phenolic contents as well as evaluating the mutagenic properties of, 12 medicinal plants used by the Vha-Venda people against venereal and related diseases. An attempt was also made toward isolating and identification of the most active compounds from some extracts that were active against Neisseria gonorrhoeae. Twelve medicinal plants and various plant parts, Adansonia digitata (bark), Acacia karroo (bark), Aloe chabaudii (roots), Bolusanthus speciosus (leaves, bark and stem), Ekebergia capensis (leaves and bark), Elephantorrhiza burkei (roots), Grewia occidentalis (roots), Osyris lanceolata (roots), Pappea capensis (leaves), Peltophorum africanum (bark), Pterocarpus angolensis (leaves and bark) and Ximenia caffra (leaves and roots) were evaluated for their antimicrobial properties against two Gram-positive (Bacillus subtilis and Staphylococcus aureus), three Gram-negative (Neisseria gonorrhoeae, Escherichia coli and Klebsiella pneumonia) bacteria and the fungus Candida albicans. The plant materials were extracted with petroleum ether (PE), dichloromethane (DCM), 80% ethanol (EtOH) and water. Methanol was used for extracting materials for phenolic contents and HIV-1RT assays. The Disc diffusion method was used to determine gonococcal percentage inhibition and a microdilution assay was used to determine minimum inhibition concentration (MIC) and minimum fungicidal concentrations (MFC). Bolusanthus speciosus and X. caffra extracts exhibited the best antigonococcal, antifungal and antibacterial activities whilst A. digitata and A. chabaudii showed poor activities. The medicinal plants were also evaluated for cyclooxygenase (COX-1 and -2) and HIV-1 reverse transcriptase inhibition activity. The DCM and PE extracts of A. digitata bark, B. speciosus bark, P. angolensis bark and P. capensis leaves showed good anti-inflammatory activity against both COX-1 and COX-2. Methanol and water extracts of B. speciosus stems, P. africanum bark, P. angolensis leaves and P. capensis leaves exhibited good anti-HIV-1 RT activity. A. chabaudii roots, E. capensis bark and O. lanceolata roots showed low HIV-1 RT percentage inhibition. Phytochemical analysis using spectrophotometric methods revealed the presence of a variety of phenolic compounds in all the plant extracts including total phenolics, flavonoids, gallotannins and condensed tannins. High levels of total phenolics, flavonoids, gallotannins and condensed tannins were detected in X. caffra. Low amounts of flavonoids, gallotannins and condensed tannins were detected in B. speciosus. The Ames test using Salmonella typhimurium tester strain TA98 with and without S9 metabolic activation revealed that all plant extracts were non-mutagenic toward S. typhimurium strains TA98 without metabolic activation. However, E. burkei roots and E. capensis bark showed mutagenic effects toward TA98 after metabolic activation. Therefore, these two plants need to be used with caution, however more studies are required to confirm this result. Good antimicrobial activity observed in X. caffra leaves prompted an attempt to isolate active compounds. A pure compound from X. caffra leaves exhibited moderate activity (63%) against N. gonorrhoeae. However, the structure of the compound has as yet to be ratified. Pharmacological activity of the twelve medicinal plants used by Vha-Venda people against venereal and related diseases were validated in this study. The results obtained in this study give credence to the use of some of these plants. This study has further confirmed the need for screening these medicinal plants for more pharmacological activities. These plants may offer a new source of chemicals for the effective treatment of venereal and related diseases. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2012.

The Orchidaceae makes up the largest and most diverse family of flowering plants. Orchids are popular, often expensive ornamentals, with a broad range of ethnobotanical applications. There is very limited documented information on South African medicinal orchid species; no formal pharmacopoeia outlining ethnobotanical uses; and ethnobotanical and distribution records are either scarce or inconsistent and plant populations are becoming gradually smaller. There have been significant developments in medicinal orchid research worldwide with medicinal use and corresponding pharmacological and phytochemical properties being extensively investigated. It is evident from the literature that there is no pharmacological research on South African medicinal orchids; hence the need to explore biological activity and chemical composition of South African medicinal orchid species. The ethnobotanical approach used to select the orchid species for pharmacological and phytochemical research elsewhere, yielded valuable biological compounds. Thus, a similar approach was applied to South African medicinal orchids. There are approximately 20 000 species and 796 genera of orchids distributed across the world. In southern Africa, orchids are widely represented with 55 genera and 494 species. Approximately 75% are endemic to this region. As part of the current investigation a review of available ethnobotanical literature on South African medicinal orchids was prepared. The review revealed that an estimated 49 indigenous orchid species from 20 orchid genera are currently being informally traded and used in South African traditional medicine. They are used primarily for medicinal and cultural purposes, especially by the Zulu community in South Africa. Medicinal uses of orchid species include: treatment of inflammatory, intestinal, neurological and reproductive disorders and emetics are used to cause emesis. Non-medicinal uses of orchid species include: love, fertility, protective and lethal charms. Based on their ethnobotanical uses and endemism, South African orchids were considered to be one of the untapped sources of bioactive compounds that needed to be researched. The current investigation addressed the broader aims of medicinal plant research by determining the efficacy, safety and chemical profile of seven indigenous orchid species used in South African traditional medicine and practices. The biological and toxic effects of orchid plant extracts were assessed using established pharmacological bioassays. The phytochemical evaluation of the seven orchid plant extracts provided insight into the classes of chemical compounds present and their possible role in the observed biological activities. The potential of plant extracts from seven orchid species used in South African traditional medicine, as sources of natural bioactive products, are discussed. The current investigation determined the biological activity and chemical profile of seven orchid species commonly traded in KwaZulu-Natal herbal markets: Ansellia africana Lindl., Bulbophyllum scaberulum (Rolfe) Bolus, Cyrtorchis arcuata (Lindl.) Schltr., Eulophia hereroensis Schltr., Eulophia petersii (Rchb.f.) Rchb.f., Polystachya pubescens (Lindl.) Rchb.f. and Tridactyle tridentata (Harv.) Schltr. Well established in vitro micro-dilution bioassays were used to determine the antibacterial, antifungal, anthelmintic activities of crude orchid extracts. A minimum inhibitory and/or lethal effect of organic and aqueous crude orchid extracts was observed against Bacillus subtilis, Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Candida albicans and Caenorhabditis elegans. Tridactyle tridentata aqueous root extract produced the most effective antibacterial activity against S. aureus (0.049 mg/ml). All T. tridentata organic root extracts produced significant inhibitory activities against B. subtilis and S. aureus. Eulophia petersii DCM pseudobulb extracts significantly inhibited all bacterial strains tested (0.39 mg/ml against S. aureus and 0.78 mg/ml against B. subtilis, E. coli, and K. pneumoniae). Eulophia hereroensis 80% EtOH root extract was the only other extract to exhibit significant inhibitory effects against K. pneumoniae (0.65 mg/ml). After 48 h C. albicans was most susceptible to P. pubescens aqueous pseudobulb extract (0.0816 mg/ml). Eulophia petersii DCM pseudobulb extract however, exhibited significant activity against C. albicans (0.65 mg/ml) over 72 h. Cyrtorchis arcuata leaf and root extracts were the most effective anthelmintic extracts with MLCs of 0.041 mg/ml for 80% EtOH leaf and root extracts; 0.049 mg/ml for aqueous leaf extracts and 0.78 mg/ml for aqueous and DCM root extracts. Caenorhabditis elegans was most susceptible to all A. africana and T. tridentata organic root extracts. A similar significant effect was observed for all E. petersii organic pseudobulb extracts, DCM extracts and organic root extracts of B. scaberulum. Only the DCM tuber and root extracts of E. hereroensis exhibited lethal effects on C. elegans. All of the P. pubescens extracts showed poor anthelmintic activity. Similarly, in vitro enzyme based cyclooxygenase (COX) 1 and 2 and acetylcholinesterase (AChE) inhibitory bioassays, revealed significant inhibition of COX-1, COX-2 and AChE enzymes by crude organic and certain aqueous orchid extracts. Out of a total of 53 evaluated extracts, 21 and 13 extracts exhibited significant anti-inflammatory activity in the COX-1 and COX-2 assays respectively. The DCM tuber extract of E. hereroensis was the only extract to significantly inhibit both COX enzymes, 100.02±0.11% and 87.97±8.38% respectively. All B. scaberulum root extracts (DCM, EtOH and water) exhibited COX-2 selective inhibitory activity (100.06±0.01, 93.31±2.33 and 58.09±3.25%). Overall, the DCM root extract of A. africana was found to be the most potent extract (EC50 0.25±0.10 mg/ml). The 80% EtOH root extract of B. scaberulum was the most potent in the COX-2 assay (EC50 0.44±0.32 mg/ml). Generally the root extracts exhibited greater AChE inhibitory activity; where the most active extract was B. scaberulum DCM root extract (EC50 0.02±0.00 mg/ml). All aqueous extracts, except that of A. africana roots and B. scaberulum pseudobulbs, showed poor or no COX-1 and COX-2 inhibition. The antioxidant capacity of crude orchid extracts was determined using: hydrogen atom transfer (HAT) (β-carotene/linoleic acid assay) and single electron transfer (SET) (2,2‟-diphenylpicrylhydrazyl (DPPH) free radical scavenging assay and ferric reducing antioxidant power (FRAP) assay) reaction-based assays. Potent antioxidant effects were observed for certain crude methanolic orchid extracts. Generally, there was a dose-dependent change in radical scavenging activities of crude extracts from which EC50 values were determined. The root extracts of all species, except that of E. petersii, had consistently more effective radical scavenging activity than that of other plant parts within each species. The pseudobulb extract of E. petersii, was the most potent extract (EC50 1.32±0.86 mg/ml). In the β-carotene-linoleic acid assay, based on the oxidation rate ratio (ORR), the leaf extract of T. tridentata and the root extracts of C. arcuata and E. hereroensis exhibited the best antioxidant effects (0.02, 0.023 and -0.15 respectively). Similarly, the average antioxidant activity (%ANT) of these samples was greater than that of BHT (95.88±6.90%) and all other samples. Bulbophyllum scaberulum leaf, pseudobulb and root extracts, E. petersii pseudobulb extract and T. tridentata root extract also exhibited a greater capacity to prevent β-carotene oxidation when compared to BHT. All crude orchid extracts tested demonstrated a general dose-dependent response in the ferric reducing power assay. The reducing power of ascorbic acid (0.08 mM) and BHT (0.05 mM), as measured as absorbance, was 1.12±0.12 and 0.73±0.08 respectively. At 6.25 mg/ml, A. africana root and E. petersii pseudobulb extracts were the most effective in reducing power activity. The short-term bacterial reverse mutation Ames Salmonella/microsome mutagenicity (ASMM) assay, which makes use of mutant histidine-dependent Salmonella typhimurium strains, was used to determine the mutagenicity and toxicity of crude orchid extracts. In the presence of a mutagen S. typhimurium TA98 strain detects frameshift events while the TA100 and TA102 strains detect base-pair substitutions. In the absence of metabolic activation, mutagenic extracts were observed against the TA98 strain only. All A. africana DCM leaf and stem extracts tested, the DCM root extract (0.5, 0.05 mg/ml) and EtOH leaf, stem and root extracts at 5 mg/ml exhibited mutagenic effects. The EtOH root extracts (5, 0.5 mg/ml) of B. scaberulum exhibited mutagenic indices (MI) comparable to that of 4NQO (17.00 and 13.00, respectively). Eulophia petersii PE pseudobulb extract demonstrated mutagenic potential at 5 mg/ml. The ethanolic root extracts of T. tridentata showed mutagenic effects at 5 and 0.5 mg/ml. The mutagenicity index (MI) with metabolic activation (S9) was determined using only the TA98 strain; where no mutagenic effects were observed. In the phytochemical evaluation of crude methanol orchid extracts, the Folin-Ciocalteu assay for total phenolics, butanol-HCl assay for condensed tannins, rhodanine assay for gallotannins and vanillin assay for flavonoids revealed a quantitative chemical profile of the tested samples. The correlation between observed biological effects and chemical compounds present was found to be generally significant. The significant antimicrobial, anthelmintic, anti-inflammatory and antioxidant activity of E. petersii pseudobulb extracts and E. hereroensis tuber and root extracts may be attributed to their high total phenolic content. Alternatively, the significant levels of gallotannin content in E. hereroensis may have contributed to the bioactivity. The flavonoid content of B. scaberulum and T. tridentata may explain the potent activity observed in the anti-inflammatory, antioxidant and acetylcholinesterase inhibitory assays; while the flavonoid content C. arcuata may have contributed to the potent anthelmintic and antioxidant activities. The significantly higher levels of gallotannin content may explain the significant anti-inflammatory and anthelmintic activity of A. africana. A number of biologically active compounds have been isolated from certain Orchidaceae species around the world on the basis of their traditional medicinal uses. The traditional uses of these orchid species were scientifically validated. No pharmacological research has been previously conducted on South African medicinal orchids; therefore the current investigation has produced novel findings on the efficacy and safety of these orchid species and promotes the continued research of medicinal orchids in South Africa. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2012.

Tulbaghia (Alliaceae) is a genus of plants with medicinal, ornamental and nutritive value. Different plant parts such as roots, bulbs, leaves and flowers are used in the treatment of a variety of conditions. The bulbs of Tulbaghia violacea are used as a remedy for pulmonary tuberculosis as well as an anthelmintic. Due to the extensive harvesting of plants in this genus, the genus is susceptible to overexploitation and may eventually become threatened with extinction. It was therefore the aim of this study to systematically examine the micropropagation of Tulbaghia ludwigiana and Tulbaghia violacea, as well as to evaluate the antimicrobial and phytochemical properties of micropropagated plants. Seeds of T. ludwigiana and T. violacea were successfully decontaminated using 70% ethanol, 1% Benlate and 3.5% NaOCl. Temperature played a significant role in the germination of both species while light did not play a significant role in this process. Light did not play a significant role in the stomatal density of T. violacea seedlings. Hypocotyls were the regenerative part of the seedlings in both species. A low number of shoots was yielded by the combination of various concentrations of NAA and mTR in the growth medium in both species. There were more isoprenoid cytokinins than there were aromatic cytokinins in each of the seedling sections of T. violacea that were analysed, with tZ being the predominant isoprenoid cytokinin, while BA was the predominant aromatic cytokinin. Shoots of both species were successfully rooted in a medium with IBA, while some shoots had simultaneously rooted during shoot multiplication. Potting soil and vermiculite were used in the acclimatization of both species, where T. violacea plantlets acclimatized successfully. This was not the case for T. ludwigiana. Micropropagated T. violacea plantlets contained higher concentrations of phytochemical compounds and displayed better antibacterial activity than outdoor-grown plants. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2011.

Livestock in southern Brazil is sustained largely by native pastures, which have large species diversity. This area is part of the Pampa biome. However, invasive species Eragrostis plana (tough lovegrass) threat to native grasslands of this biome. Becomes necessary to develop alternative methods of control of this species, since conventional control practices, although fairly efficient, has no as a principle the preservation of native vegetation. The present work aimed to study the control of tough lovegrass through biocontrol method for isolation of potentially pathogenic fungi, as well as the allelopathic effects of aqueous extract of carqueja (Baccharis trimera) on the germination of tough lovegrass. The fungi were isolated from seeds of tough lovegrass and incubated for eight days of growth at 25 °C and a photoperiod of 12 hours, then identified to genus chamber. Five genera were identified: Alternaria sp., Curvularia sp., Aspergillus sp., Penicillium sp. e Fusarium sp. To test the interaction of these fungi with the seeds of E. plana, fungi inoculation was performed in seeds through deposition technique, which were previously sterilized with 1% NaClO, and seeded on two types of substrates: filter paper (in vitro experiment) and substrate and sand mixture (ex vitro experiment). Treatments consisted of the five identified fungi, inoculated separately in seeds of E. plana. The genera showed that the best results were: Fusarium sp., Curvularia sp. e Penicillium sp. The carqueja belongs to the Asteraceae family, possessing compounds with many properties. Among the compounds present in B. trimera can highlight the presence of flavonoids, tannins, fatty acids, steroids, triterpenoids. To evaluate the allelopathic effect of B. trimera on E. plana different doses of the crude aqueous extract (CAE) of carqueja were applied on the seeds of tough love-grass. The concentrations where 0; 25; 50; 75 and 100 g.L-1. The experiments were conducted in vitro and ex vitro. The High-performance liquid chromatography (HPLC) was also conducted to identify the compounds present in the CAE. Were found in larger amounts: caffeic acid, chlorogenic acid, ellagic acid and isoquercitina. In the in vitro experiment the inhibition of the germination appeared from the concentration of 25 g.L-1 upwards, causing a diminishing germination with increasingly higher concentrations of the CAE. In the ex vitro experiment the reduction of emergence was lower, but was statistically significant. The data show that the carqueja extracts interfere on the development of the tough lovegrass by reducing leaf length and the dry mass of the aerial part and of the roots. / A pecuária na Região Sul do Brasil é sustentada, em grande parte, por pastagens nativas, que apresentam grande diversidade de espécies. Esse território faz parte do bioma Pampa. A espécie invasora capim-annoni ameaça essas pastagens. Torna-se necessário o desenvolvimento de métodos alternativos de controle dessa espécie, pois práticas convencionais de controle, embora relativamente eficientes, não tem a preservação da vegetação nativa como princípio. O presente trabalho teve por objetivo estudar o controle de capim-annoni, através do biocontrole por método de isolamento de fungos potencialmente fitopatogênicos, bem como dos efeitos alelopáticos do extrato aquoso de carqueja (Baccharis trimera) sobre a germinação do capim-annoni. Os fungos foram isolados a partir de sementes de E. plana e incubados por oito dias em câmara de crescimento, a 25 °C e fotoperíodo de 12 horas, em seguida identificados até gênero. Foram identificados cinco gêneros: Alternaria sp., Curvularia sp., Aspergillus sp., Penicillium sp. e Fusarium sp. Para testar a interação desses fungos com as sementes de E. plana, foi realizada a inoculação dos fungos nas sementes através da técnica de deposição, sendo elas previamente desinfestadas com NaClO 1%, e semeadas em dois tipos de substratos: papel-filtro (experimento in vitro) e mistura de substrato e areia (experimento ex vitro). Os tratamentos constaram dos cinco fungos identificados, inoculados separadamente nas sementes de E. plana. Os gêneros que apresentaram os melhores resultados foram Fusarium sp., Curvularia sp. e Penicillium sp. A carqueja, pertencente à família Asteraceae, apresenta vários compostos na sua composição. Dentre os compostos presentes em B. trimera pode-se destacar a presença de flavonóides, taninos, ácidos graxos, esteróides, triterpenóides. Para avaliar o efeito alelopático de B. trimera em E. plana foram aplicadas diferentes doses de extrato bruto aquoso (EBA) de carqueja nas sementes de capim-annoni. As concentrações foram 0; 25; 50; 75 e 100 g.L-1. Os experimentos foram conduzidos in vitro e ex vitro. Também foi realizada a técnica Cromatografia Líquida de Alta Eficiência (HPLC) para identificar os compostos presentes no EBA. Foram encontrados em maior quantidade: ácido cafeico, ácido clorogênico, isoquercitina e ácido elágico. No experimento in vitro a inibição da germinação deu-se a partir da concentração 25 g.L-1, sendo que a germinação diminuiu conforme a concentração do EBA aumentou. No experimento ex vitro a redução da emergência de plântulas foi estatisticamente significativa. Os dados obtidos mostram que os extratos de carqueja interferem no desenvolvimento do capim-annoni para comprimento de folhas e massa seca de parte aérea e raiz.

Os produtos naturais constituem uma das fontes mais promissoras para o descobrimento de novos ativos na terapêutica do câncer. O carcinoma de laringe é um dos mais comuns tipos de câncer envolvendo as áreas de cabeça e pescoço, e apresenta elevada taxa de morbidade e mortalidade em pacientes com estágio avançado. Terapias alternativas e/ou adjuvantes para o tratamento deste tipo de câncer representam uma necessidade emergente. Uma das alternativas mais promissoras é o desenvolvimento de nanocarreadores contendo ativos antitumorais. Araucaria angustifolia (Bert. O Kuntze) pertence à família Araucariaceae e é uma planta reconhecidamente medicinal. Seus estróbilos femininos dão origem a pinha, constituída por pinhões (sementes verdadeiras) e brácteas (sementes não desenvolvidas). O objetivo do presente estudo foi avaliar o efeito antiproliferativo do extrato aquoso de brácteas de A. angustifolia (EAA) em células tumorais de laringe HEp-2 e seus mecanismos de ação. Além disso, a possibilidade de associar o EAA a nanoesferas (NE) a fim de potencializar seu efeito antitumoral também foi investigado. A análise química por meio de Espectrometria de Massas de Alta Resolução evidenciou a presença majoritária de polifenóis no EAA. Os resultados mostraram que o EAA induziu citotoxicidade nas células tumorais HEp-2 através de dois diferentes ensaios de viabilidade celular (ensaio de MTT e de exclusão do corante Trypan blue). No entanto, o mesmo não foi capaz de induzir citotoxicidade significativa em células normais HEK-293, utilizadas como controle, indicando um efeito diferencial seletivo do EAA sobre as células tumorais. As células HEp-2 tratadas com EAA apresentaram níveis aumentados de peroxidação lipídica, danos oxidativos a proteínas e aumento da produção de ON, juntamente com depleção das defesas antioxidantes superóxido dismutase (Sod) e catalase (Cat). Além disso, o EAA induziu danos ao DNA, juntamente com fragmentação nuclear e condensação da cromatina nestas células. Alterações nos marcadores epigenéticos, como hipometilação do DNA e redução da atividade de DNMT1 foram também observadas. A exposição das células tumorais ao extrato aumentou a expressão de proteínas apoptóticas de via intrínseca mitocondrial, mediada pela ativação da proteína Bax, liberação de AIF e foi independente da ativação de p53. O EAA modificou o metabolismo energético das células tumorais, elevando os níveis de piruvato desidrogenase (PDH) e estimulando a fosforilação oxidativa mitocondrial. Embora tenha ativado a mitocôndria destas células, o mesmo causou falhas no potencial de membrana mitocondrial (ΔΨm), juntamente com diminuição dos níveis de proteínas dos complexos I e III da CTE, inibição da atividade do complexo I, produção de ERO e depleção de ATP. A associação do EAA à NE permitiu a obtenção de sistemas com tamanho de partícula inferior a 200 nm, índice de polidispersibilidade abaixo de 1, potencial zeta negativo e pH estável pelo período de 30 dias, na condição testada. No entanto, a taxa de associação obtida foi baixa (19%), indicando a necessidade de futuros estudos a fim de aumentar a eficiência de incorporação do extrato. As NE per se demonstaram capacidade de reduzir a viabilidade de células tumorais e induzir alterações redox, sensibilizando estas células, demonstrando ser um possível carreador para a vetorização de ativos antitumorais. O conjunto de dados deste estudo demonstra a potencialidade dos compostos presentes nas brácteas de A. angustifolia para o desenvolvimento de novas estratégicas terapêuticas para o câncer. / Submitted by Ana Guimarães Pereira (agpereir@ucs.br) on 2016-12-19T15:24:14Z No. of bitstreams: 1 Tese Cátia dos Santos Branco.pdf: 11116932 bytes, checksum: 02a668c3fc8a7376e3031fe7b838d152 (MD5) / Made available in DSpace on 2016-12-19T15:24:14Z (GMT). No. of bitstreams: 1 Tese Cátia dos Santos Branco.pdf: 11116932 bytes, checksum: 02a668c3fc8a7376e3031fe7b838d152 (MD5) Previous issue date: 2016-12-19 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, CAPES. / Natural products are among one of the most promising fields in finding new active substances in cancer therapy. Laryngeal carcinoma is one of the most common cancers affecting the head and neck regions, and is associated with high morbidity and mortality in patients with the advanced stage. Alternative and/or complementary therapies for treating this cancer represents an emerging need. One of the most promising alternatives is the development of nanocarriers containing antitumor substances. Araucaria angustifolia (Bert. O Kuntze) belongs to Araucariaceae family and it is recognized as medicinal plant. Female strobilus originates the pinecone, which contains seeds and undeveloped seeds, commonly known as bracts. The aim of this study was to evaluate the antiproliferative effects of A. angustifolia bracts aqueous extract (AAE) in HEp-2 cancer cells and its action mechanisms. Moreover, the possibility to associate AAE in nanospheres (NS) to improve its antitumor effect was also investigated. Chemical analysis using High Resolution Mass Spectrometry (HRMS) revealed the major presence of polyphenols in AAE. The results showed that AAE induced cytotoxicity in HEp-2 cells, by using two differents approaches (MTT and Trypan blue assays). However, the extract was not able to induce significant cytotoxicity in HEK-293 normal cells used as control, indicating a selective differential effect of AEE in tumor cells. HEp-2 treated cells presented high levels of lipid peroxidation, oxidative damage to proteins and increment on NO production, along with depletion on antioxidante defenses superoxide dismutase (Sod) and catalase (Cat). In addition, AAE induced DNA damage, nuclear fragmentation and chromatin condensation in these cells. Epigenetic alterations, such as DNA hypomethylation and DNMT1 activity were also observed. Cell exposition to the AAE increased expression of proteins of the mitochondrial intrinsic pathway, via Baxtriggered, along with AIF release, and it is independent of p53 incitement. AAE changed energetic metabolism of cancer cells, increasing levels of pyruvate dehydrogenase (PDH) and stimulating mitochondrial oxidative phosphorylation. Although it enabled the mitochondria of these cells, the extract caused loss of mitochondrial membrane potential (ΔΨm), reduction on protein expression levels of complex I and III, inhibition of complex I activity, ROS generation and ATP depletion. The association of EAA to NS allowed obtaining systems with particle size lower than 200 nm, polydispersity index less than 1, negative zeta potential and pH stable for the period of 30 days, under tested condition. However, the association rate obtained was low (19%); therefore, further studies are needed to improve AAE encapsulation. The NS per se demonstrated ability to reduce the viability of tumor cells and induce redox stress, sensitizing these cells, proving to be a possible carrier for delivery of antitumor substances. The data set of this study demonstrates the potentiality of the compounds present in the A. angustifolia bracts for the development of new therapeutic strategies for cancer.

Made available in DSpace on 2015-05-14T13:00:01Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 1293356 bytes, checksum: eb48e7877c4f4bdbd473d0f996411c16 (MD5) Previous issue date: 2014-07-31 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / There are some strains of Escherichia coli that are pathogenic and often cause urinary tract infections, septicemia and meningitis in neonates. Some produce enzymes known as β-lactamases of extended spectrum (ESBL) that decrease the therapeutic options, thereby arousing interest in the discovery of new antibacterial products. Medicinal plants and their essential oils are especially rich in metabolites with antimicrobial properties as soon researches are developed in searches of these substances. Thus, the present study aimed to evaluate the effect of essential oil of Coriandrum sativum and its major phytochemicals against strains of E. coli ESBL. Was initially determined the sensitivity of the tested strains to conventional antibiotics, as well as screening of the antibacterial activity of the oils against these strains. Chosen C. sativum oil, its chemical composition was determined by gas chromatography coupled to mass spectrometer (GC/MS) and the antibacterial activity of both the oil as the major phytochemicals were evaluated by determination of Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC), the microdilution technique, the kinetics of microbial death oil was also analyzed. It has also played a modulating effect over the conventional antibiotics. It was found that the tested strains are producing ESBL. Among the phytochemicals, linalool (39.78%) presented itself as a major component, followed by linalool oxide (27.33%). The oil and linalool inhibited all strains having MIC and MBC of 256 μg/mL, 512 μg/mL and 1024 μg/mL, between 512 and 1024 μg/mL, respectively. Linalool oxide was inactive at the maximum test concentration of 1024 μg/mL. This oil has antibacterial activity dependent on its concentration and time of exposure in the micro-organism. Both oil and linalool modulates the action of the antibiotics ciprofloxacin and cefoxitin, synergistic interaction of oil and ciprofloxacin being observed. The combination with linalool showed synergistic effects with ciprofloxacin and norfloxacin. The results suggest that the essential oil of C. sativum can suppress the growth of species E. coli ESBL and its major phytochemical despite having weak antibacterial activity, may modulate the action of some antibiotics. / Existem algumas cepas de Escherichia coli que são patogênicas e frequentemente causam infecções urinárias, septicemias e meningites em neonatos. Algumas produzem enzimas conhecidas como β-lactamases de espectro estendido (ESBL) que diminuem as opções terapêuticas, despertando assim o interesse pela descoberta de novos produtos antibacterianos. As plantas medicinais e principalmente seus óleos essenciais são ricos em metabólitos com propriedades antimicrobianas, logo pesquisas são desenvolvidas em buscas destas substâncias. Assim, o presente estudo teve como objetivo avaliar o efeito do óleo essencial de Coriandrum sativum e seus fitoconstituintes majoritários contra cepas de E. coli ESBL. Inicialmente foi determinado o perfil de sensibilidade das cepas ensaiadas a antibióticos convencionais, bem como a triagem da atividade antibacteriana dos óleos contra estas cepas. Escolhido o óleo de C. sativum, a sua composição química foi determinada por Cromatografia Gasosa acoplada a Espectrômetro de Massa (CG/EM) e a atividade antibacteriana tanto do óleo quanto dos fitoconstituintes majoritários foram avaliadas pela determinação da Concentração Inibitória Mínima (CIM) e Concentração Bactericida Mínima (CBM), pela técnica de microdiluição, a cinética de morte microbiana do óleo também foi analisada. Verificou-se também o efeito modulador desempenhado sobre os antibióticos convencionais. Constatou-se que as cepas ensaiadas são produtoras de ESBL. Entre os fitoconstituintes, o linalol (39,78%) apresentou-se como principal componente, seguido pelo óxido de linalol (27,33%). O óleo e o linalol inibiram todas as cepas apresentando CIM e CBM de, 256 μg/mL, 512 μg/mL e 1024 μg/mL, entre 512 e 1024 μg/mL, respectivamente. O óxido de linalol não apresentou atividade na concentração máxima ensaiada de 1024 μg/mL. Este óleo possui atividade antibacteriana dependente de sua concentração e do tempo de exposição no micro-organismo. Tanto o óleo quanto o linalol modulou a ação dos antibióticos ciprofloxacina e cefoxitina, sendo observada interação sinérgica do óleo e a ciprofloxacina.Já a combinação com linalol, apresentou efeitos sinérgicos com a ciprofloxacina e norfloxacina. Os resultados sugerem que o óleo essencial de C. sativum pode suprimir o crescimento de espécies de E. coli ESBL e que seu fitoconstituinte majoritário apesar de ter atividade antibacteriana fraca, pode modular a ação de alguns antibióticos

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / Syzygium cumini is a plant originated from India and adapted in Brazil, popularly known as black plum, jambolan and jamun. In this study, the antioxidant and protective activities of ethanolic leaves extract of Syzygium cumini (EE) were evaluated on paracetamol-induced liver damage in mice. The total antioxidant activity was determined by ABTS and DPPH methods and the content of vitamin C, extractable polyphenols, anthocyanins, yellows flavonoids and carotenoids were evaluated by different methods. The toxicity of EE was evaluated with different doses up to 1200 mg/kg administered orally for eight consecutive days. Subsequently, doses of the EE (150, 300 and 600 mg/kg weight) were administered by gavage for 7 consecutive days before treatment with single dose of paracetamol (500 mg/kg, orally) and its hepatoprotective effect has been studied by monitoring biochemical parameters. The extract presented high antioxidant activity in ABTS (2148.88  140.26 mM Trolox/g of sample) and DPPH (87.11  1.58 g of sample/g DPPH) assays. The extractable polyphenols prevailed among determined phytochemicals. Regarding toxicity, EE showed no toxic effects at the doses evaluated. EE (600 mg/kg) presented hepatoprotective effect that was evidenced by the decrease in the activities of alanine aminotransferase (56.94%), aspartate aminotransferase (30.14%) and alkaline phosphatase (32.68%) in the serum of animals treated with EE (600 mg/kg) and subjected to ingestion of paracetamol (EE600+ P group) compared with the group paracetamol. In liver tissue, there was a decrease of 25.68%, 31.67% and 31.97% of the malondialdehyde levels and elevation of 60.03%, 63.67% and 69.12% of the dismutase superoxide activity in EE150+ P EE300+ P and EE600+ P groups, respectively, when compared with the paracetamol group. Furthermore, non-protein sulfhydryls groups in the liver increased 56.74% and 63.61% in EE300+P and EE600 + P groups when compared with the paracetamol group. The ethanolic leaves extract of Syzygium cumini conferred hepatoprotection against stress by paracetamol, this effect may be associated to its high antioxidant activity. / Syzygium cumini à uma planta originÃria da Ãndia e adaptada ao Brasil, popularmente conhecida como azeitona da terra, ameixa preta, jambolÃo e jamun. Neste estudo, as atividades antioxidantes e protetoras do extrato etanÃlico (EE) de folhas de azeitona da terra foram avaliadas contra danos induzidos por paracetamol no fÃgado de camundongos. A atividade antioxidante total foi determinada atravÃs dos mÃtodos ABTS e DPPH e os teores de vitamina C, polifenois extraÃveis, antocianinas, flavonoides amarelos e carotenoides foram avaliados por diferentes mÃtodos. A toxicidade do EE foi avaliada com diferentes doses atà 1200 mg/Kg administradas por via oral durante oito dias consecutivos. Posteriormente, doses do EE (150, 300 e 600 mg/Kg de peso) foram administradas por gavage durante 7 dias consecutivos antes do tratamento com dose Ãnica de paracetamol (500 mg/Kg, por via oral) e o seu efeito hepatoprotetor foi estudado atravÃs do monitoramento de parÃmetros bioquÃmicos. O extrato apresentou elevada atividade antioxidante nos ensaios de ABTS (2.148,88  140,26 ÂM Trolox/g de amostra) e DPPH (87,11  1,58 g de amostra/g DPPH). Os polifenois extraÃveis predominaram entre os fitoquÃmicos determinados. Em relaÃÃo à toxicidade, EE nÃo apresentou efeitos tÃxicos nas doses avaliadas. EE (600 mg/kg) apresentou efeito hepatoprotetor que foi evidenciado pela diminuiÃÃo nas atividades de alanina aminotransferase (56,94%), aspartato aminotransferase (30,14%) e fosfatase alcalina (32,68%) no soro de animais tratados com EE (600 mg/kg) e submetidos a ingestÃo de paracetamol (grupo EE600+P) em comparaÃÃo com o grupo paracetamol. No tecido hepÃtico, houve diminuiÃÃo de 25,68%, 31,67% e 31,97% dos teores de malondialdeÃdo e elevaÃÃo de 60,03%, 63,67% e 69,12% da atividade da superÃxido dismutase nos grupos EE150+P, EE300+P e EE600+P, respectivamente, quando comparados com o grupo paracetamol. Ademais, os grupos sulfidrilas nÃo proteicos no fÃgado aumentaram 56,74% e 63,61% nos grupos EE300+P e EE600+P quando comparados com o grupo paracetamol. O extrato etanÃlico de folhas de azeitona da terra conferiu hepatoproteÃÃo contra estresse por paracetamol, podendo esse efeito estar associado à sua elevada atividade antioxidante.

Submitted by (edna.saturno@ufrpe.br) on 2016-07-26T11:34:06Z No. of bitstreams: 1 Aldenise Chagas Curvello G Moreira.pdf: 1832678 bytes, checksum: 08dcbab15cec31bc62f7c570ddae3ae1 (MD5) / Made available in DSpace on 2016-07-26T11:34:06Z (GMT). No. of bitstreams: 1 Aldenise Chagas Curvello G Moreira.pdf: 1832678 bytes, checksum: 08dcbab15cec31bc62f7c570ddae3ae1 (MD5) Previous issue date: 2011-02-24 / The aim this work was to characterize "cajá-umbu" fruits coming from the germplasm's collection IPA (Instituto Agronômico de Pernambuco). Fruits of the 4 genotypes (G 2.2, G 2.3, G 3.1, G 3.2) were used to quantify the phytochemicals bioactive and evaluate the antioxidant potential. The polyphenols were quantified by spectrophotometric method, the ascorbic acid by titulometric method and carotenoids by chromatographic and spectrophotometric techniques. Hydroacetone, hydromethanolic and aqueous extracts were obtained using 3 extraction systems: a) sequential extraction: acetone 80%, methanol 80% and water, b) sequential extraction: water, methanol 80% and acetone 80% c) non¬sequential extraction. The isolated and combined extracts were submitted to the determination of total phenolic content, screened for their free DPPH- (1,1¬diphenyl-2-picrilhidrazil) and ABTS-+) (2,2'-azino-bis-(3-etilbenzotiazoline 6¬sulfonic acid) scavenging activity, and their effect on the linoleic acid peroxidation by the system of co-oxidation r3-carotene/acid linoleic and ferric thiocyanate method. The fruits showed significant content of polyphenols, low levels of ascorbic acid and carotenoids. The I3-carotene 5-6, 5'6'-diepoxide was the major component of genotypes G 2.2, G 2.3, G 3.1, and 5-6 epoxide lutein for G 3.2. The extraction procedure which employed acetone 80% as first solvent could extract a larger amount of total phenolic. Ali genotypes exhibited good DPPH- scavenging activity (ECSD value of 0.30 to 0.43, TEcSD value of 8.98 to 15.24 minutes, and EA value of 0.18 to 0.40), significant ABTS-+ scavenging capacity (55.56 a 120.08 IJMol TEAC.g-1) and high percentage of inhibition of peroxidation of linoleic acid (88.45 to 92.89%). In co-oxidation of linoleic acid/r3-carotene system exhibited moderate activity (56.91 to 69.41 % inhibition), showing that they are effective in blocking the onset of oxidation reaction (F1 <1), but the propagation phase, G 3.2 genotype showed higher efficiency (F2 <1). Therefore, fruits of "cajá-umbuzeiro" have bioactive compounds and good antioxidant potential that can contribute te the dietary intake of antioxidant. / Com o objetivo de caracterizar frutos de genótipos de cajá-umbuzeiras cultivados no Banco de Germoplasma do IPA (Instituto Agronômico de Pernambuco), frutos de 4 genótipos (G 2.2; G 2.3; G 3.1; G 3.2) foram utilizados para quantificar principais fitoquímicos bioativos e avaliar o potencial antioxidante. Os polifenóis foram quantificados por método espectrofotométrico; o ácido ascórbico por método titulométrico e os carotenóides por técnicas• cromatográfica espectrofotométrica. Os extratos hidroacetônico, hidrometanólico e aquoso forar obtidos empregando-se 03 sistemas de extração: a) extração sequencial: acetom a 80%, metanol a 80% e água; b) extração sequencial: água, metanol a 80% e acetona a 80%; c) extração não sequencial. Os extratos isolados e combinados foram avaliados quanto ao teor de fenólicos totais, a capacidade de sequestrar o radical 1, 1-difenil-2-picrilhidrazil (DPPHe) e o radical 2,2' -azino-bis-(3-etilbenzotiazolina-6-ácido sulfônico (ABTSe+), a inibição da oxidação em sistema da co-oxidação l3-caroteno/ácido linoléico e pelo método tiocianato férrico. Os frutos apresentaram teor relevante de polifenó!s, baixos teores de ácido ascórbico e de carotenóides. O l3-caroteno-5,6-5'6'-diepóxido foi o componente majoritário dos genótipos G 2.2; G 2.3; G 3.1, e a luteína-5,6 epóxido do G 3.2. O procedimento de extração que empregou a acetona 80% como primeiro solvente conseguiu extrair maior quantidade de fenólicos totais. Todos os genótipos exibiram relevante capacidade de sequestro do radical DPPH (ECsD de 0,30 a 0,43, T ECSD de 8,98 a 15,24 minutos, e EA de 0,18 a 0,40), expressiva capacidade de sequestro do radical ABTSe+ (55,56 a 120,08 IlMol TEAC.g-1) e forte inibição da peroxidação do ácido linoléico (88,45 a 92,89%). Em sistema da co-oxidação do ~-caroteno/ácido linoléico exibiram ação moderada (56,91 a 69,41 % de inibição), demonstrando serem eficientes em bloquear o início da reação) de oxidação (F1 <1), porém na fase de propagação, o genótipo G 3.2 demonstrou maior eficiência (F2<1). Os frutos do cajá-umbuzeiro apresentam em sua constituição compostos bioativos e bom potencial antioxidante, podendo contribuir com o aporte dietético de antioxidante.

Made available in DSpace on 2014-08-20T13:42:04Z (GMT). No. of bitstreams: 1 Dissertacao_Milene_Teixeira_Barcia.pdf: 1315431 bytes, checksum: 076c98a13af9fd70d5553f28c620d0ff (MD5) Previous issue date: 2009-09-01 / The jambolanum is a tree originating from India and Indonesia, belonging to the family of Mirtaceas. The tree bears fruit during January to May in several Brazilian states. The fruit are small, purpleand involves a single core. The taste of jambolanum not stand out so much because the astringency of the fruit. The consumption of tropical fruit is growing in the last years due to the appeal of nutritional and therapeutic effects. These fruits contain different phytochemicals, many of these exhibit antioxidant properties, especially the tocopherols, carotenoids and phenolic compounds, because its property of reacting with free radicals. Among the phenolic compounds there are the flavonoids that chemically, include anthocyanins and flavonols. Practically do not know the chemical composition of jambolanum because; therefore, the knowledge of some of its components is important to predict the feasibility of their use as in natura foods and as raw material for the production of other products. This study aimed to evaluate the chemical composition of the fruits of jambolanum collected from three cities in the southern state of RS. The experiment consists of three trees in three different cities of the southern of RS(Santa Vitória do Palmar, Pelotas and Capão of Leão), and its was analyzed for chemical and phytochemical compositiont.By the results it was found that the jambolanum fruit shows the chemical composition similar to other fruits, presenting low amount of ash, proteins, pectin and fiber. The moistureand sugar content and pH is characteristic of citrus fruit; however, the fruit showed higher soluble solids content. The Jambolanum fruit presents low tocopherols and ascorbic acid content, but it showed a high content of tannins and phenol compounds. / O jambolão é uma árvore originária da Índia e Indonésia, pertencente à família das Mirtáceas. Frutifica nos meses de janeiro a maio, em diversos estados do Brasil. Os frutos são pequenos, de coloração roxa e envolve um caroço único. O sabor do jambolão não se destaca muito devido a adstringência da fruta. O consumo de frutas tropicais vem aumentando nos últimos anos devido ao apelo pelo seu valor nutritivo e aos efeitos terapêuticos. Estas frutas contêm diferentes fitoquímicos, muitos destes exibem propriedades antioxidantes, com destaque para os tocoferóis, carotenóides e compostos fenólicos, devido suas propriedades de reagir com os radicais livres presentes. Dentre os compostos fenólicos destacam-se os flavonóides que quimicamente, englobam as antocianinas e os flavonóis. Praticamente não se conhece a composição química do jambolão, devido a isto, o conhecimento de alguns de seus componentes se faz importante, para prever a viabilidade de sua utilização como alimento in natura e como matéria prima para a elaboração de outros produtos. Este estudo apresentou como objetivo avaliar a composição química das frutas de jambolão colhidas de diferentes plantas de três cidades da Região Sul do estado do RS. O experimento consta de frutas oriundas de três árvores distintas em três cidades do Sul do estado (Santa Vitória do Palmar, Pelotas e Capão do Leão), sendo avaliadas quanto a composição química e ao teor de fitoquímicos. Pelos resultados observa-se que o jambolão apresenta composição química similar a outras frutas características da região, apresentando baixo conteúdo em cinzas, proteínas, pectina e de fibras. O teor de umidade, açúcares e pH é característico de frutas cítricas, no entanto, com um teor superior em sólidos solúveis. A fruta apresenta baixo conteúdo em tocoferóis e ácido ascórbico, porém apresenta um alto teor de taninos e de compostos fenólicos.

Made available in DSpace on 2014-08-20T13:42:04Z (GMT). No. of bitstreams: 1 Dissertacao_Andressa_Carolina_Jacques.pdf: 729903 bytes, checksum: c29b8dc4bad8abf286333f2238a71dc8 (MD5) Previous issue date: 2009-07-31 / Brazil is considered one of the major consumer´s country of fruits, occupying the third worldwide position. The diversity of fruit for the market is growing, but their properties and activities are not fully determined. However, from the beginning of the 90s, a large fruit supply is justifying studies related to the development of new products, which in most cases concentrates the major consumption form as raw fruit or pulp. This work aimed to produce pulp from blackberry cv. Tupy, and store it under different temperature conditions (-10, -18 and -80 ºC) for 6 months, evaluating the stability of its main phytochemicals after processing and during the storage period. The results showed that the temperature of -10 °C was not sufficient to cause significant changes in relation to total phenols, total anthocyanins, antioxidant capacity and titrable acidity (2 months). The temperature of -18 ° C was not sufficient to cause significant changes in the pulp with respect to: total phenols and antioxidant capacity (4 months), total anthocyanins (2 months) and β-carotene (6 months). At temperature of -80 ° C, few changes were caused in the pulp stored for 6 months, and observed only small changes in soluble solids, hydroxybenzoic acid. Total and individual carotenoids in any of the three IPF temperatures sufficient to avoid losses. In Tocopherol, only in the pulp stored at -80 ° C for 2 months there was no change.The ascorbic acid was completely degraded within 6 months of storage at all temperatures (-10, -18 and -80 °C). The phenolics were the compounds that most contributed to the antioxidant capacity of blackberry cv. Tupy. / O Brasil é considerado um dos principais paises consumidores de frutas, ocupando a terceira posição mundial. A diversidade de frutas destinadas ao mercado é cada vez maior, mas suas propriedades e atividades não estão totalmente determinadas. No entanto, a partir do início da década de 90, a grande oferta de frutas vem justificando estudos direcionados ao desenvolvimento de novos produtos, que, na maioria das vezes, concentra na fruta in natura e na polpa suas maiores formas de consumo. Este trabalho teve por objetivo elaborar polpa de amora-preta da cultivar Tupy, e armazená-la sob diferentes condições de temperaturas (-10, -18 e -80 ºC) durante 6 meses, e avaliar a estabilidade de seus principais fitoquímicos após o processamento e durante o período de armazenamento. Os resultados demonstraram que a temperatura de -10°C foi suficiente para não causar alterações significativas com relação à fenóis totais, antocianinas totais, capacidade antioxidante e acidez titulável (2 meses). A temperatura de -18°C foi suficiente para não causar alterações significativas nas polpas com relação à: fenóis totais e capacidade antioxidante (4 meses), antocianinas totais (2 meses) e β-caroteno (6 meses). Na temperatura de -80°C, poucas foram as alterações causadas na polpa armazenada por 6 meses, sendo observado apenas pequenas alterações em sólidos solúveis, ácido hidroxibenzóico. Nos carotenóides totais e individuais nenhuma das três temperaturas fpi suficiente para evitar as perdas. Nos tocoferois, apenas na polpa armazenada a -80°C por 2 meses não houve alteração. E o ácido ascórbico foi totalmente degradado nos 6 meses de armazenamento nas duas temperaturas (-10 e -18°C). Os compostos que mais contribuíram para o poder antioxidante da amorapreta cv. Tupy foram os compostos fenólicos.