Dissertations / Theses on the topic 'Medicinal plants (features pharmacology)'

Campomanesia xanthocarpa Berg, espécie pertencente à família Myrtaceae, é conhecida popularmente como gabiroba. Diversas propriedades terapêuticas são atribuídas às espécies de Campomanesia, tais como, combate à disenteria, febre, cistites e uretrites. O presente trabalho objetiva contribuir para elaboração de monografia da espécie, através do estudo químico e farmacológico do extrato hidroalcoólico liofilizado e da droga vegetal constituída de folhas. Os caracteres farmacobotânicos da droga vegetal são apresentados com o intuito de auxiliar a diagnose. Dentre as principais características macroscópicas das folhas desidratadas podem ser citadas: ondulação nas bordas; pontos translúcidos na lâmina, tipo de venação, forma da lâmina e dos pecíolos, e odor aromático. As características anatômicas que se destacaram são: mesofilo dorsiventral; grandes idioblastos contendo cristal prismático na região paliçádica; predominância de estômatos anomocíticos nas folhas hipoestomáticas; cavidades secretoras globosas, associadas a ambas as superfícies, recobertas por células dispostas aos pares em que a parede comissural mostra-se reta, sinuosa ou de \"zig-zag\"; feixe vascular bicolateral e sistema disposto em arco aberto; cristais prismáticos na região floemática. Fotomicrografias ilustraram o trabalho. A triagem fitoquímica da droga vegetal e do extrato liofilizado (EHA) indicou a presença de flavonóides, taninos, saponinas e óleo essencial. O teor de óleo essencial nas folhas frescas, foi de 0,11 %. Linalol (29%) e globulol (20%) foram identificados como os principais componentes do óleo. O teor de taninos na droga foi de 2,86 % e no extrato EHA, de 8,49 %. O teor de saponinas na droga foi de 6,27 % e no extrato EHA de 16%. O extrato apresentou elevada atividade antioxidante no modelo de medida de produção de malonildialdeído, com Q1/2 =0,2891 µg/mL. No ensaio da inibição da lipoperoxidação, induzida por sulfato de cobre, o extrato EHA reduziu significativamente o tempo de indução (\"Lag-time\") e o tempo de terminação (\"Peak-time\") da oxidabilidade de lipoproteínas de baixa densidade (LDL). O extrato EHA apresentou atividade antiúlcera no modelo de indução aguda por ácido clorídrico em etanol, com percentual de proteção de 62%. O extrato EHA apresentou atividade antimicrobiana com: CMI >1.000 e <500 µg/mL em relação a Staphylococcus aureus; CMI <500 e >100 µg/mL para Salmonella cholerasuis e CMI <1.000 e >500 µg/mL em relação a Candida albicans. O extrato EHA mostrou atividade citotóxica no ensaio da letalidade de artêmias, com DL50 de 0,503 mg/mL. O extrato EHA não apresentou toxicidade no ensaio de toxicidade aguda, na dose de 5g/kg de peso do animal, por via oral.
Campomanesia xanthocarpa Berg, a species that belongs to the Myrtaceae family, is popularly known as gabiroba. Several therapeutic properties are attributed to the various Campomanesia species, such as treating diarrhea, fever, cystitis and urethritis. This project aims at contributing through a chemical and pharmacological study of lyophilized hydrated alcohol extract and the vegetable drug made from leaves. The pharmacological and botanical features of the vegetable drug are indicated in order to help with the diagnosis. Main macroscopic features of the dehydrated leaves include: wavy edges, translucent blade spots, venation type, blade and leafstalk forms and scent. The main anatomical features are: dorsiventral mesophyll; large idoblasts containing prismatic crystal in the palisade parenchyma; predominance of anomocytic stomata in the hypostomatic leaves, globose segregating cavity associated to both surfaces, covered by cells organized in pairs where the commissure wall appears straight, sinuous, or in zigzag; bicollateral vascular bundle and system organized in an open arch, prismatic crystals in the phloem region. Photomicographs illustrate the study. The phytochemical screening of the vegetable drug and the lyophilized extract (EHA) indicated the presence of essential oil, flavonoids, tannins and saponins. The essential oil content in the fresh leaves was 0.11 %. Linalol (29%) and globulol (20%) were identified as the main oil components. Tannin content was 2.86% in the drug and 8.49% in the EHA extract. The saponin content was 6.27% in the drug and 16% in the EHA extract. The extract displayed a high antioxidant activity in the model of malonyl dialdehyde production measure with Q1/2 = 0,2891 µg/mL. In the copper sulfate-induced lipoperoxydation inhibition assay the EHA extract significantly reduced Lag-time and Peak-time for low-density lipoprotein (LDL) oxidability. The EHA extract displayed antiulceration activity in the acute induction model by hydrochloric acid in ethanol, with a 62% protection percentage. The EHA extract displayed antibacterial activity with: CMI >1,000 and <500 µg/mL relative to Staphylococcus aureus; CMI <500 and >100 µg/mL for Salmonella cholerasuis and CMI <1.000 and >500 µg/mL regarding Candida albicans. The EHA extract displayed cytotoxic activity in the artemias lethality trial, with DL50 of 0,503 mg/mL. The EHA extract displayed no toxicity in the acute toxicity trial in the 5g/kilo oral dosage per animal body weight.

This thesis describes phytochemical and biological/pharmacological studies on six medicinal plants. Plant materials were extracted by ethanol, tested for their biological activity and then subjected to "bioassay-guided fractionation" to yield active and inactive compounds. The compounds were identified by standard physico-chemical techniques including UV, IR, NMR and ElMS. The crude ethanol extract of the leaves of Premna shcimperi was found to be antibacterial against gram positive bacteria (Staphylococcus aureus and Bacillus subtilis). An antibacterial bioassay-guided fractionation of the extract resuited in the isolation of the active principle, (5R,8R,9S,10R)-12-oxo-ent-3,13(16)- clerodien-15-oic acid (SHM-l). This novel compound was bactericidal against S. aureus and B. subtilis at concentrations of 25 and 50 µg ml⁻¹. Studies on the structural-activity relationship of SHM-l showed that the α,β unsaturated moiety played a major role in its antibacterial activity. Investigation of the inactive fractions of P. schimperi extract afforded three flavonoid aglycones: luteolin, quercetin and kaempferidej three flavonoid glycosides: luteolin-4" -,β-D-glucoside, quercetin-3-,β-D-galactoside and quercetin-α-L-arabinopyranoside and five cinnamate and benzoate derivatives. The ethanol extract of the leaves of Premna oligotricha was also found to be antibacterial against a range of gram-positive bacteria. A bioassay-directed isolation afforded three novel active principles, two diterpenes: 16-hydroxy-clerod- 3,13(14}-diene-15,16-olide (SHM-3) and ent-12-oxolabda-8,13(16}-dien-15-oic acid (SHM-S) and a sesquiterpene, 7-α, hydroxy-2-oxo-6,11-cyclofarnes-3(15)-ene (SHM-19). While the activity of SHM-5 and SHM-19 was marginal that of SHM-3 was good, almost comparable with streptomycin. The antimicrobially inactive fractions of P. oligotricha extract yielded a novel diterpene, ent-8,β12α-epidioxy- 12,β-hydroxlabda-9(11), 13-dien-15-oic acid γ lactone and two novel flavonoids: 3,5,5'-trihydroxy-6,7 ,3' ,4'-tetramethoxyflavone and 3,5,7 ,5'-tetrahydroxy-6,3',4'- trimethoxyflavone. Investigation of the leaves of Premna recinosa afforded six flavonoids: quercetin, luteolin, pachypodol, chrysosplenol-D, naringenin and eriodictyol and three lignans: {+)-8-hydroxy-pinoresinol, {+)-lariciresinol and {-)-seco-isolariciresinol. Portulaca oleracea and Pentas schimperiana are used in the folk medicine as local anaesthetics. Studies on the isolated nerve and/or muscle preparations showed that extracts do block nerve conduction in these excitable tissues. A subsequent systematic bioassay-guided fractionation of crude extracts showed that excess potassioum ions in the extracts were responsible for the in vitro pharmacological activity of P. oleracea and P. schimperiana. Extracts from the leaves of Leonotis ocymifolia var raineriana showed no antibacterial activity. Three novel diterpenes: ent- {13S)-9,13α-epoxylabda- 6{ 19),β,16{15)-diol dilactone, (13R)-6,β-acetoxy-9, 13α-epoxylabda-19(20),8,16{ 15)- diol dilactone and 20,8-acetoxy-9α,13e-dihydroxy-15(16)-epoxy-Iabd-14-en-6,19,(β- O-lactone together with a known compound: {13S)-20,β-acetoxy-9,13α-epoxylabda- 6(19),β,16{15)-diol dilactone were isolated. Investigation of the root afforded two known compounds 3-methoxy-4-hydroxy-{trans)-cinnamaldehyde and 3-methoxy- 4-hydroxy-bezaldehyde.

This thesis describes investigations of the usage of unconventional therapeutic methods to treat diabetes mellitus, with particular reference to Asian patients. Findings suggest that usage of unconventional therapeutic methods may persist in diabetic patients regardless of their language, religious belief, ethnic and cultural background or psychological states and adherence. The thesis is presented in two parts. Study 1, a preliminary study, was conducted in Thailand. Groups of adults with diabetes mellitus, aged 17 - 70+ years, were studied to assess the extent to which unconventional therapeutic methods were used, and to examine the possibility that such usage is associated with their psychological states and unsatisfactorily feelings toward orthodox medicine. Data collection was achieved through a combination of well-established and well-evaluated questionnaires and a structured interview. A scale to assess attitudes to diabetes was found to be reliable in this sample, but scales to measure diabetes knowledge and treatment satisfaction were not. Study 2 was a study of British Indo-Asians in Foleshill, Coventry, England. The extent to which medicinal plants were used was explored and compared between two different cultural and religious backgrounds of adults with diabetes: (1) born in an Asian country and (2) born in England. The majority of participants were old with low educational background and income. A number of modifications were made to the structured interview used in study 1 to make it more appropriate for this sample. The two studies suggest that usage of medicinal plants is common among diabetics in Thailand and among British Indo-Asian diabetics born in an Asian country. Only a minority of users of medicinal plants in both countries were willing to discuss their usage of medicinal plants with their physicians. This could be because users believed that their physicians might not approve the usage of non-orthodox treatment. In Thailand, usage of medicinal plants was significantly associated with one factor - a lack of basic diabetes knowledge. In the study in England, a typical user was characterised as an Asian female born in an Asian country, who had a low income, used betel-nut, had a preferencef or a doctor's ethnicity, and had low treatment satisfaction and adherence scores.

Early man dabbled with the use of plant extracts to cure ailments. This practice has been passed down from generation to generation and today more than 50% of the world'sdrugs are natural products or derivatives thereof. Scientists have thus established a branch of research called natural product research. This branch of research involves the identification and purification of secondary metabolites with a specific biological activity. The methodology involves the screening of plant products for a specific biological activity, purification of the biologically active natural product by separation technology and structure determination. The biologically active natural products is then further scrutinized to serve as a novel drug or lead compound for the development of a novel drug. This research exploited this research methodology.

>Magister Scientiae - MSc
Research into traditional medicines is often conducted in a multidisciplinary approach as motivated by a desire to understand them in as complete a manner as possible,realizing their chemistry, biology and pharmacology. One biological approach involves monitoring the cytotoxicity of the extracts of subfractions against the nauplii,Artemia salina (brine shrimp). Organic and aqueous extracts of seven South African medicinal plants was investigated for biological activity. Selected plant extracts was also evaluated for AChE inhibitory activity. The objectives of this study was to look for any correlation between known biological activities of the investigated plants and BSLT lethality data and also to look for any correlation between AChEI activity and BSLT lethality data for selected plant extracts. The most active of the plants was the n-hex extract of T.alliacea, followed by the aqueous extract of C.mellei and the MeOH extract of C.quadrifidus; the MeOH and the DCM extracts of A.afra; the DCM extract of P.undulatum and the EtOAc extract of A.annua. The results from this study show a good correlation with antitumor, antimicrobial and anti-trypanocidal activity.The various plants extracts investigated showed good inhibitory activity towards AChE using the TLC bioautography method. The results obtained from this study indicate that this type of activity is not only subject to plants containing alkaloids, but rather a diverse class of compounds may exhibit this kind of activity. The extracts that showed good AChE inhibitory activity also showed good cytotoxicity towards brine shrimp nauplii.

Bibliography: p. 128-140.
The term "traditional medicine" refers to the ways of protecting and restoring health that existed before the arrival of modern medicine. These approaches to health belong to the traditions of each country and have been handed down from generatio to generation.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior
Protium heptaphyllum March (Burseraceae) populary known as âalmÃcegaâ is a popular medicinal plant largely encountered in the Amazon region, various States of Brazil and in several South American Countries. The oily amorphous exudate obtained from this plant is widely used in skin diseases, healing of ulcers, and as an analgesic and anti-inflamatory agent. Phytochemical studies reveled the presence of several monoterpenes and pentacyclic triterpenes such as a mixture of α- e β â amyrin, maniladilol and breine. The present study aimed to investigate the general toxicity and to establish the pharmacological activity of resin and is major triterpenoid mixture, the α and β â amyrin. In toxicity tests, both the resin and triterpene mixture exhibited low toxicity to mice. Resin at doses up 5 g/kg, (p.o.) or 2 g/kg (i.p.) and triterpene mixture up to 3 g/kg, (p.o.) or 2 g/kg (i.p.) failed to induce any mortality in mice. In Artemia-lethality test, the calculated (probit analysis) CL50 values for resin and triterpenes were in the order of 42,54 Â 19,96 and 400 Â 27,85 μg/mL, respectively. In pharmacological tests, the resin was analysed for anti-inflamatory (carrageenan-induced edema, cotton pellet-induced granuloma, and vascular permeability increase induced by i.p. acetic acid) and gastroprotective (absolute ethanol and acidified ethanol) effects, whereas the triterpene mixture was examined in assays that demostrate gastroprotective (against lesions induced by absolute ethanol), antipruritus (against pruritus induced by Dextran T40 and compound 48/80), acute and visceral antinociceptive (test of subplantar and intracolonic capsaicin) and hepatoprotective (against acetaminophen- and Ga1N/LPS-induced models of hepatitis) effects. In anti-inflammatory test, the resin (200 e 400 mg/kg, p.o.) although failed to modify the carrageenan-induced acute rat paw-edema response, it caused signficant inhibitions at a dose of 400mg/kg on the formation of cotton pellet-induced granulomas and on the vascular permeability increase induced by i.p. acetic acid in mice. In addition, the (200 e 400 mg/kg) showed gastroprotective potential against absolute- and acidified ethanol- induced gastric lesions as evidenced from siginificant diminution in lesion scores, restoration of the ethanol-induced depletion of non-protein sulfhydryl content More over, the resin demonstrated an antisecretory effect on gastric acid secretion induced in 4-h pylorus ligated rats. The triterpene mixture also produced similar gastroprotection against ethanol-induced lesions in a manner similar to capsaicin, a pungent principle from hot peppers. This protection possibly involves capsaicin-sencitive primary afferents since it was abolished in mice pretreated with a neurotoxic dose of capsaicin. The α and β â amyrin mixture (100 mg/kg) manifested antipruritus effect as evidenced from suppression of scratching behaviour in the mouse model of prurits induced by s.c. injections of dextran T40 and compound 48/80. Besides, it also produced an antiedematogenic effect in model of hind paw edema induced by histamine, compound 48/80 and dextran T40 and markedly depressed the compound 48/80-elicited rat mast cell degranulation (ex vivo). An antinociceptive effect of triterpenoid mixture (3-100 mg/kg) was observed in capsaicin-evoked somatic (1.6 μg/site, suplantar) and visceral (149 μg, intracolonic) models of nociception in mice. Greater suppression of nociceptive behaviors were evidenced at a dose of 10 mg/kg α and β â amyrin mixture, which mimicked the effect produced by ruthenium red, a non-competitive capsaicin antagonist. The antinociceptive effect of triterpenoid mixture was found to be naloxone (2 mg/kg)- sensitive, suggesting an opioid mechanism. A blockade by triterpene mixture was also evidenced on the hyperthermic but not the hypothermic response of subcutaneously administered capsaicin (10 mg/kg) suggesting possible incolvement of TRPV1 receptor. In open-field and rota-rod tests, the triterpene mixture did not manifest signs of either sedation or motor abnormality in mice that could account for the observed antinociception. In the model of acetaminophen (500 mg/kg)-induced hepatotoxicity, the triterpenoid mixture (50 and 100 mg/kg) effectively reduced the elevated serum AST and ALT levels, restored the depleted GSH and markedly diminished the histopathological alterations. Potentation of pentobarbital-sleeping time was, however observed at these doses of triterpenoid, incidating a probable suppression of cytochrome P450 and thus a diminished metabolite formation that may account for reduced acetaminophen toxicity. The α- and β â amyrin mixture offered complete protection against the mortality associated with Ga1N/LPS , but caused only a moderate diminution of serum enzymes and histopathological alterations. Taken together, these findings show that the resin and α- and β â amyrin mixture possess low toxicity and have a wide therapeutic potential with anti-inflammatory, antinociceptive, antipruritus, and gastro- and hepato-protective actions. Most of the effects of triterpenoid mixture appear to involve in part the participation of primary sensory afferents in their actions.
A espÃcie Protium heptaphyllum (Aubl.) March (Burseraceae) popularmente conhecida como almÃcega, à encontrada na regiÃo AmazÃnica, em vÃrios Estados do Brasil e paÃses da AmÃrica do Sul. Esta espÃcie exsuda uma resina oleosa e amorfa, usada na medicina popular como analgÃsico, cicatrizante e expectorante. Estudos fitoquÃmicos demonstraram a presenÃa de monoterpenos e triterpenos pentacÃclicos, tais como α - amirina e β - amirina, maniladilol e breina. O presente trabalho teve como objetivo investigar os efeitos tÃxicos e farmacolÃgicos da resina e de seus constituintes majoritÃrios, a mistura de triterpenos α e β â amirina. Na avaliaÃÃo dos efeitos tÃxicos observamos a toxicidade aguda destes produtos em camundongos e Artemia sp. Analisando os efeitos sistÃmicos, avaliamos a atividade antiinflamatÃria da resina (edema de pata induzido por carragenina, granuloma induzido por âpelletsâ de algodÃo e permeabilidade vascular induzida por Ãcido acÃtico) e da mistura de α e β â amirina (edema induzido por histamina, serotonina, dextrana T40 e composto 48/80). Examinamos ainda as atividades gastroprotetora e antisecretÃria da resina (lesÃes gÃstricas induzidas pelo etanol absoluto e etanol acidificado e secreÃÃo Ãcida induzida pela ligaÃÃo pilÃrica) e as atividades gastroprotetora (lesÃes gÃstricas induzidas pelo etanol absoluto, com animais dessensibilizados por capsaicina), antipruriginosa (prurido induzido pelo dextrana T40 e composto 448/80 e desgranulaÃÃo de mastÃcitos ex vivo) antinociceptiva (nocicepÃÃo induzida pela administraÃÃo subplantar e intracolÃnica de capsaicina, resposta hipotÃrmica induzida por capsaicina) e hepatoprotetora (lesÃes hepÃticas induzidas por acetaminofeno e Ga1N/LPS) da mistura de α e β â amirinas. NÃo foi possÃvel estabelecer as DL50 da resina (atà 5 g/kg, v.o. e 1 g/kg, i.p.) e da mistura de α e β â amirina (atà 3 g/kg, v.o. e atà 2 g/kg, i.p.) em camundongos. A mistura de α e β â amirina, mas nÃo a resina, mostrou toxicidade para Artemisa sp, sendo as CL50 de 42,54  19,96 e 400  27,85 μg/mL, respectivamente. Nos modelos de permeabilidade vascular induzido por Ãcido acÃtico (camundongo) e granuloma induzido por âpelletâ de algodÃo (ratos), a resina demonstrou efeito antiinflamatÃrio significativo na dose de 400mg/kg, reduzindo a permeabilidade vascular e o peso seco do granuloma. Contudo, a reina nÃo apresentou atividade sobre edema induzido por carragenina (ratos). Adicionalmente, a resina preveniu as lesÃes gÃstricas induzidas por etanol absoluto e etanol acidificado, alÃm de impedir a depleÃÃo dos grupos sulfidrilas produzida pelo etanol absoluto nas doses de 200 e 400 mg/kg. Um efeito antisecretÃrio da resina (200 e 400mg/kg) foi observado no modelo de secreÃÃo Ãcida induzida pela ligaÃÃo pilÃrica em ratos. A mistura de α e β â amirina tambÃm exibiu atividade gastroprotetora inibindo as lesÃes gÃstricas por etanol absoluto, cujo mecanismo parece envolver os neurÃnios sensoriais primÃrios sensÃveis à capsaicina. A administraÃÃo oral dos triterpenos α e β â amirina (100 mg/kg), apresentou atividade antiedematogÃnica, nos modelos de edema de pata induzidos por histamina, composto 48/80 e dextrana T40, mas nÃo sobre o edema induzido por serotonina. A atividade antipruriginosa tambÃm foi observada com as α e β â amirina nas doses variando de 50 a 200 mg/kg, em modelos de prurido induzido por dextrana T40 e pelo composto 48/80 e na reduÃÃo (100 mg/kg) da degranulaÃÃo de mastÃcitos peritoneais ex vivo pelo composto 48/80. O efeito antinociceptivo da mistura, nas doses de 3 a 100 mg/kg, foi verificado atravÃs da inibiÃÃo dos comportamentos de nocicepÃÃo induzidos pela administraÃÃo subplantar ou intracolÃnica de capsaicina em camundongos. A antinocicepÃÃo produzida por estes triterpenos (10 mg/kg) sobre o tempo de lambedura induzido pela capsaicina (1,6 μg/20 μL) nÃo foi potencializada nem revestida pelo vermelho de rutÃnio (1,5 mg/kg), mas foi significativamente inibida pela naloxona (2 mg/kg), sugerindo mecanismo opiÃide. A participaÃÃo dos receptores α2 - adrenÃrgicos neste efeito tambÃm foi eliminada, tendo em vista que a ioimbina nÃo reverteu o efeito antinociceptivo das amirinas no modelo de nocicepÃÃo visceral induzida pela capsaicina. Estes triterpenos bloquearam ainda a hipertermia induzida pela capsaicina (10 mg/kg), mas nÃo reverteram a resposta hipotÃrmica induzida por este agente, sugerindo a participaÃÃo do receptor vanilÃide (TRPV1) no efeito antinociceptivo das amirinas. Nos modelos de hepatoxidade, a mistura de α e β â amirina (50 e 100 mg/kg) reduziu o aumento dos nÃveis sÃricos de ALT e AST e restabeleceu os nÃveis de GSH hepÃticos, diminuindo as alteraÃÃes histopatolÃgicas induzidas pelo acetaminofeno (500 mg/kg), alÃm de potencializar o tempo de sono induzido por pentobarbital sÃdico (50 mg/kg), indicando que este efeito hepatoprotetor envolve a inibiÃÃo do citocromo P â 450. A mistura ofereceu ainda completa proteÃÃo contra a mortalidade induzida por Ga1N/LPS, reduzindo as lesÃes hepÃticas em camundongos e reduzindo os nÃveis sÃricos de ALT, mas nÃo de AST ou GSH hepÃticos, sugerindo um possÃvel feito neuroimunomodulatÃrio neste modelo. Os triterpenos α e β â amirina nas doses variando de 3 a 30 mg/kg, nÃo manifestam efeitos sedativos ou incoordenaÃÃo motora em camundongos. A resina e mistura de α e β â amirina possuem baixa toxicidade e atividades antiinflamatÃria e gastroprotetora. Os triterpenos α e β â amirina exibiram atividade antipruriginosa, antinociceptiva e hepatoprotetora, cujos efeitos envolvem, pelo menos em parte, a participaÃÃo dos neurÃnios aferentes sensoriais primÃrios.

Panax ginseng C.A. Mey. (Araliaceae), which has been known in China for more than 4000 years, occupies a particular place among tonic remedies. Pharmacological investigations of the roots have shown that the basic effect of ginseng's action is its capacity to increase the nonspecific resistance of the organism to various untoward influences. In addition to ginseng, another plant of the Araliaceae family is used in a similar manner and is known as Siberian ginseng, Eleutherococcus senticosus (Rupr. et Maxim.) Maxim.). Its roots have been very extensively investigated by Russian scientists. Ginseng is reported to be shorter in duration and weaker in activity when compared to E. semicosus in respect to stimulant and tonic effects. 13 Chinese scientists have claimed that China Produced E. senticosus can exert a definite anti fatigue action. Less extensive studies have been reported on the berries of Schisandra chinensis (Turcz.) Baill. (Magnoliaceae). for its adaptogenic properties. The main objective of the work reported in the following pages was to design and test a simpler and easier animal test model whereby multiple physiologic parameters such as Tn EKG and respiratory rates could be continuously and concurrently monitored. It was desired that the test model could be used repeatedly and rapidly to serve as a screen for all potential adaptogenic plants and their derivatives and extracts.

The use of medicinal plants is present since the ancient times of civilization, currently being used as a complement in the treatment of pathologies. In Sergipe, more specifically in Mussuca community in Orange, the use of medicinal plants is closely linked to the cultural aspects of the community. Medicinal plants in Mussuca can be found in large quantities in the backyards of residents. Even as this massive amount of medicinal plants from various genres, few studies of pharmacological prospecting etnodirigida performed in maroon territories. In this sense, this work was to carry out an ethnopharmacological survey of medicinal plants with antinociceptive properties in maroon community Mussuca and is presented in three chapters. In the first chapter described the general introduction, objectives and literature review. In the second chapter an ethnobotanical survey was presented from popular knowledge for the identification of medicinal plants used in the antinociceptive activity Mussuca community in pain management. And in the third chapter a pharmacological study was conducted to identify the antinociceptive effect of the six plants indicated primary use by local experts. The ethnobotanical data were collected through participant observation and semi-structured interviews. In parallel, we applied the snow ball method in which it was possible to identify seven local experts, where each expert indicated an analgesic plant primary use. The quantitative ethnobotanical data were analyzed using the technique use value and the qualitative results were analyzed using the technique collective subject discourse. We conclude that the experts of the village have empirical knowledge about medicinal plants and technical analgesic use value helped identify the species of greatest significance to the community. For pharmacological testing of the writhing test was performed to identify the antinociceptive effect of plant species by reducing the writhing. The results were analyzed by 1-way ANOVA followed by Bonferroni test. The program used was Graph Pad Prism version 4.0. It was found that all the medicinal plants has antinociceptive effect, with the more potent the Schinus terebinthifolius Raddi, more effective Rolandra fruticosa (L.) Kuntze, Guarea guidonia, Cecropia pachystachya Trécul, Schinus terebinthifolius Raddi.
O uso de plantas medicinais está presente desde os tempos imemoriais da civilização, sendo utilizadas atualmente como complemento no tratamento de patologias. Em Sergipe, mais especificadamente na comunidade Mussuca em Laranjeiras, o uso de plantas medicinais está intimamente ligado aos aspectos culturais da comunidade. As plantas medicinais na Mussuca podem ser encontradas em grande quantidade nos quintais das casas dos moradores. Mesmo como essa quantidade maciça de plantas medicinais de diversos gêneros, poucos são os trabalhos de prospecção farmacológica etnodirigida realizados em territórios quilombolas. Neste sentido, esse trabalho consistiu em realizar um levantamento etnofarmacológico de plantas medicinais com propriedades antinociceptivas na comunidade quilombola Mussuca e está apresentado em três capítulos. No primeiro capítulo foi descrito a introdução geral, objetivos e revisão de literatura. No segundo capítulo foi apresentado um levantamento etnobotânico a partir do conhecimento popular para a identificação de plantas medicinais com atividade antinociceptiva utilizadas na comunidade Mussuca no manejo da dor. E no terceiro capítulo foi realizado um estudo farmacológico visando identificar o efeito antinociceptivo das seis plantas de uso principal indicada pelos especialistas locais. Os dados etnobotânicos foram levantados por meio da observação participante e entrevistas semiestruturadas. Paralelamente foi aplicado o método bola de neve em que foi possível identificar sete especialistas locais, onde cada especialista indicou uma planta analgésica de uso principal. Os resultados etnobotânicos quantitativos foram analisados através da técnica valor de uso e os resultados qualitativos foram analisados através da técnica discurso do sujeito coletivo. Conclui-se que os especialistas do povoado possuem conhecimento empírico sobre plantas medicinais analgésicas e a técnica valor de uso ajudou a identificar as espécies vegetais de maior significância para a comunidade. Para o teste farmacológico foi realizado o teste de contorção abdominal visando identificar o efeito antinociceptivo das espécies vegetais através da diminuição da contorção abdominal. Os resultados foram analisados através de Anova de 1 via, seguido do teste de Bonferroni. O programa utilizado foi o Graph Pad Prism versão 4.0. Constatou-se que todas as plantas medicinais possui efeito antinociceptivo, sendo a mais potente a Schinus terebinthifolius Raddi e mais eficazes as Rolandra fruticosa (L.) Kuntze, Guarea guidonia, Cecropia pachystachya Trécul e Schinus terebinthifolius Raddi.

Pela crescente utilização das plantas medicinais e óleos essenciais na prevenção e tratamento de enfermidades, embasados nas propriedades antibacterianas, antifúngicas e antioxidantes é que objetiva-se com o presente trabalho identificar a composição e atividade antioxidante dos óleos essenciais de Lavandula angustifolia, Pogostemon cablin, Rosmarinus officinalis, Thymus vulgaris, Hedyosmun brasiliense, Psidium guajava, Baccharis dracunculifolia e Schinus terebinthifolius, bem como avaliar o potencial antibacteriano in vitro frente Staphylococcus aureus; Salmonella enteretidis;Escherichia coli e Pseudomonas aeruginosa e a atividade antifúngica in vitro frente o Fusarium graminearum.Os óleos e as cromatografias foram obtidos por hidrodestilação com parcerias externas. A atividade antibacteriana foi determinada por meio da microdiluição em caldo. Enquanto que a atividade antifúngica dos óleos de Baccharis dracunculifolia e Pogostemon cablin foi determinada por solubilização dos óleos no meio de cultura e dispondo um disco miceliar de F. graminearum. A atividade antioxidante foi determinada pela ação sequestradora do radical 1-difenil-2-picrilidrazina (DPPH). Como resultados obtiveram-se as constituições dos óleos essenciais principalmente por monoterpenos como 32,5% de cânfora no óleo de Rosmarinus officinalis, 47,0% de timol no óleo de Thymus vulgaris, 31,5% de patchulol em Pogostemon cablin; e ésteres como 40,1 % de acetato de lanalina no óleo de Lavandula angustifolia. Nos óleos das plantas nativas, a composição foi basicamente de sesquiterpenos como 17,58% de cis trans_nerolidol no óleo de Baccharis dracunculifolia, 31,6% de germacreno B no Hedyosmun brasiliense; 16,1% de α-selineno no óleo de Psidium guajava e 18,6% de α-bergamoteno no de Schinus terebinthifolius. A atividade antibacteriana do óleo de Thymus vulgaris, apresentou os resultados mais efetivos, com concentrações inibitórias mínimas (CIM) e concentrações bactericidas mínimas (CBM) de 0,195 e 1,56 µl/mL frente S. aureus; 0,195 e 50 µl/mL respectivamente para S. enteritidis; para E. coli a CIM foi de 0,390 µl/mL e CBM 0,780 µl/mL e para P. aeruginosa 0,780 e 12,5 µl/mL. As atividades antifúngicas foram determinadas frente ao F. graminearum tendo o Pogostemon cablin (8,0 µl/mL), no tempo 96 h inibido 80,0% do fungo. A atividade antioxidante do óleo essencial de Pogostemon cablin foi superior aos demais, com 12,08 µmol trolox/mL-1 de atividade sequestradora do radical. Sugere-se que os óleos de Lavandula angustifólia, Thymus vulgaris, Rosmarinus officinalis, Baccharis dracunculifolia, Psidium guajava e Schinus terebinthifolius, podem ser utilizados no combate de infecções causadas por S. aureus, E. coli, S. enteritidis e mais estreitamente por P. aeruginosa. E que o óleo de Pogostemon cablin possa servir como agente antifúngico frente ao fitopatógeno F. graminearum, além do óleo de Thymus vulgaris desempenhar importante atividade antioxidante.
The growing use of medicinal plants and essential oils in order to prevent and carrie the diseases treatment, based on the antibacterial, antifungal and antioxidant properties is that the aim of this work, whichs ought to identify the composition and antioxidant activity of some essential oils as Lavandula angustifolia, Pogostemon cablin, Rosmarinus officinalis, Thymus vulgaris, Hedyosmun brasiliense, Psidium guajava, Baccharis dracunculifolia and Schinus terebinthifolius, as well as evaluate the antibacterial potential in vitro against Staphylococcus aureus; Salmonella enteretidis; Escherichia coli e Pseudomonas aeruginosa and antifungal activity in vitro againt Fusarium graminearum. The oils and chromatographies were obtained by hydrodistillation through external partnerships. While the antifungal activity of Baccharis dracunculifolia and Pogostemon cablin oils laying out a mycelial disc of F. graminearum. The scavenging activity of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical. As a result It was obtained the constitutions of essential oils mainly monoterpenes with 32.5 % of camphor in the Rosmarinus officinalis oil, 47 % of thymol in Thymus vulgaris, 31.5% of patchoulol in Pogostemon cablin; and esters with 40.1% of lanalin acetate in the Lavandula angustifolia oil. In the native plants oils the composition was basically of sesquiterpenes. Such as 17.58% of cis trans nerolidolin the Baccharis dracunculifolia oil; 31.6% of germacrene B in the lemon balm; 16.1% α-selinene in the Psidium guajava oil and 18.6% of α-bergamotene in the Schinus terebinthifolius.. The antibacterial activity of Thymus vulgaris oil has shown the most effective results, with minimal inhibitory concentration (MIC) and minimum bacterial concentration (MBC) of0.195 e 1.56 µL.mL-1 against S. aureus; and 0.195 e 50 µL.mL-1respectively for S. enteritidis; and for E. coli, the MIC was 0,390 µL.mL-1and MBC 0.780 µL.mL-1 and for P. aeruginosa 0.780 and 12.5 µL.mL- 1.The antifungal activities were determined against F. graminearum with Pogostemon cablin (8.0 μL.mL-1) during the period of 96h, It has inhibited 80.0% of the fungus. The antioxidant activity of the Pogostemon cablin essential oil was higher than the others with 12.08 µmol trolox/mL-1 radical scavenging activity. Based on this study, It is suggested that oils as Lavandula angustifólia, Thymus vulgaris, Rosmarinus officinalis, Baccharis dracunculifolia, Psidium guajava and Schinus terebinthifolius, can be used against infections caused by S. aureus, E. coli, S. enteritidis and more closely by P. aeruginosa. Also it was found out that Pogostemon cablin oil may serve as an antifungal agent against the phytopathogen F. graminearum. Moreover, the thyme oil can play an important antioxidant activity.

Conselho Nacional de Desenvolvimento Científico e Tecnológico
Pain is one of the major health problems, leading people to search for treatment. A pharmacological screening showed that the Caesalpinia pyramidalis ethanol extract (EE) reduces nociception behavior in the writhing, formalin, and hot plate tests in mice. In order to assess the mechanism of action of EE of C. pyramidalis, substrate, antagonists or inhibitors of different pathways involved in nociception were used in the writhing model. To select the dose, mice (n = 6 per group) were pretreated with EE of C. pyramidalis (10, 30, or 100 mg/kg, p.o.) vehicle (Tween 80, 0.2% in saline, 10 mL/kg, p.o.) or acetylsalicylic acid (ASA, 300 mg/kg, p.o.) 1 hour before the administration of acetic acid (0.6%, 0.1 ml/10 g, i.p.). Subsequently, different groups of animals were pretreated, intraperitoneally, with L-arginine (NO precursor, 600 mg/kg, 15 min), methilene blue (NO/cGMP, way inhibitor, 20 mg/kg, 15 min), glibenclamide (K+-ATP channels inhibitor, 3 mg/kg, 15 min), atropine (muscarinic-cholinergic receptors antagonist, 0.1 mg/kg, 15 min), prazosina (α1-adrenoceptor antagonist, 0.15 mg/kg, 15 min), yohimbine (α2-adrenoceptor antagonist, 0.15 mg/kg, 15 min), haloperidol (dopaminergic receptors antagonist, 2 mg/kg, 15 min), caffeine (adenosinergic receptors antagonist, 3 mg/kg, 15 min), flumazenil (gabaergic receptors antagonist, 3 mg/kg, 15 min) or, reserpine (inhibitor of monoamine receiving, 5 mg/kg, 24 h), and were treated with EE of C. pyramidalis (30 mg/kg, p.o.), vehicle (p.o.), L-NOARG (NOS inhibitor, 75 mg/kg, i.p.), acetylcholine (muscarinic receptor agonist, 1 mg/kg, i.p.), phenylephrine (α1-adrenoceptor agonist, 1 mg/kg, i.p.), clonidine (α2-adrenoceptor agonist, 0,1 mg/kg, i.p.), adenosine (adenosinergic receptors agonist, 100 mg/kg, i.p.), diazepam (gabaergic receptors agonist, 1,5 mg/kg, i.p.) or, clomipramine (neuronal receiving monoamines inhibitor, 10 mg/kg, i.p.). Sixty min after the different treatments writhing was conducted. Besides, another animal groups, 60 min before the administration of vehicle, doses of 10, 30, or 100 mg/kg of the EE of C. pyramidalis and 30 min before administration of the received morphine (3 mg/kg, i.p.) in order to be subjected to testing of capsaicin (20 μL, 1.6 mg/paw) or glutamate (20 μL, 20 μmol/paw). All experimental controls functioned as expected pharmacological action, observing their reversals of the analgesic effect, when comparing with animals that received different treatments with the drugs used. The number of writes was lower when received the EE from C. pyramidalis in the doses of 30 and 100 mg/kg (P < 0.001), in a dose-dependent manner, when was compared with the animals that received vehicle, choosing the dose of 30 mg/kg (p.o.) to this study. There was a reversal of the antinociceptive effect of the EE C. pyramidalis action when the animals were pretreated with L-arginine (P < 0.001), methylene blue (P < 0.05), glibenclamide (P < 0.001), atropine (P < 0.001), yohimbine (P < 0.05), and flumazenil (P < 0.001) and the reversion was not observed when administered prazosin, haloperidol, reserpine, and caffeine. The results showed, that the animals reduced the licking/biting time at the dose of 100 mg/kg in the capsaicin test (P < 0.05) and in all doses in the glutamate test (P <0.01). Concludes that the EE of C. pyramidalis presents its antinociceptive effect, acting synergistically in pathways of L-arginine/NO, NO/cGMP, K+ channel-ATP sensitive, muscarinic cholinergic, α2-adrenergic and GABAergic, and involve the participation of glutamate and capsaicin in the antinociceptive effect.
A dor representa um dos principais problemas de saúde, levando a população a uma busca constante por tratamento. Um screening farmacológico mostrou que o extrato etanólico (EE) da Caesalpinia pyramidalis reduz o comportamento de nocicepção nos testes de contorções abdominais, formalina e placa quente em camundongos. Com o objetivo de avaliar o mecanismo de ação do EE da C. pyramidalis, substrato, antagonistas ou inibidores de diferentes vias envolvidas na nocicepção foram utilizados no modelo de contorções abdominais. Para a escolha da dose, camundongos (n = 6/grupo) foram tratados por via oral com EE da C. pyramidalis (10, 30 ou 100 mg/kg), veículo (Tween 80 a 0,2% em salina, 10 mL/kg) ou ácido acetilsalicílico (AAS, 300 mg/kg) 1 hora antes da administração do ácido acético (0,6%, 0,1 mL/10 g, i.p.). Posteriormente, diferentes grupos de animais receberam pré-tratamento por via intraperitoneal com L-arginina (precursor do NO, 600 mg/kg, 15 min), azul de metileno (inibidor da via NO/GMPc, 20 mg/kg, 15 min), glibenclamida (inibidor de canais K+ATP, 3 mg/kg, 15 min), atropina (antagonista de receptores colinérgico-muscarínico, 0,1 mg/kg, 15 min), prazosina (antagonista de α1-adrenoceptores, 0,15 mg/kg, 15 min), ioimbina (antagonista de α2-adrenoceptores, 0,15 mg/kg, 15 min), haloperidol (antagonista de receptores dopaminérgicos, 2 mg/kg, 15 min), cafeína (antagonista de receptores adenosinérgicos, 3 mg/kg, 15 min), flumazenil (antagonista de receptores gabaérgicos, 3 mg/kg, 15 min) ou reserpina (inibidor da recaptação de monoaminas, 5 mg/kg, 24 h), e foram tratados com o EE da C. pyramidalis (30 mg/kg, v.o.), veículo (v.o.), L-NOARG (inibidor da NOS, 75 mg/kg, i.p.), acetilcolina (agonista de receptores muscarínicos, 1 mg/kg, i.p.), fenilefrina (agonista de α1-adrenoceptores, 1 mg/kg, i.p.), clonidina (agonista de α2-adrenoceptores, 0,1 mg/kg, i.p.), adenosina (agonista de receptores adenosinérgicos, 100 mg/kg, i.p.), diazepam (agonista de receptores gabaérgicos, 1,5 mg/kg, i.p.) ou clomipramina (inibidor da recaptação neuronal de monoaminas, 10 mg/kg, i.p.). Sessenta min após os diferentes tratamentos realizou-se o teste das contorções abdominais. Outros grupos de animais, 60 min antes da injeção da capsaicina ou glutamato, receberam a administração de veículo, doses de 10, 30 ou 100 mg/kg do EE da C. pyramidalis e, 30 min antes, receberam a administração da morfina (3 mg/kg) afim de serem submetidos aos testes da capsaicina (20 μL, 1,6 μg/pata) ou glutamato (20 μL, 20 μmol/pata). Todos os controles do experimento funcionaram conforme ação farmacológica esperada, observando-se as respectivas reversões do efeito antinociceptivo, ao se comparar os animais que receberam os diferentes tratamentos com as drogas utilizadas. O número de contorções abdominais foi significativamente menor quando os animais receberam as doses de 30 e 100 mg/kg (p < 0,001), de maneira dose-dependente, quando comparadas às observadas nos animais que receberam veículo, escolhendo-se a dose de 30 mg/kg (v.o.) para este estudo. Houve reversão do efeito antinociceptivo do EE da C. pyramidalis quando os animais receberam pré-tratamento com L-arginina (P < 0,001), azul de metileno (P < 0,05), glibenclamida (P < 0,001), atropina (P < 0,001), ioimbina (P < 0,05) e flumazenil (P < 0,001), e a reversão não foi observada quando administrados prazosina, haloperidol, reserpina e cafeína. Observou-se uma redução do tempo lambida/mordida na dose de 100 mg/kg no teste da capsaicina (P < 0,05) e em todas as doses no teste do glutamato (P < 0,01). Conclui-se que o EE da C. pyramidalis apresenta seu efeito antinociceptivo, agindo de maneira sinérgica nas vias dos sistemas L-arginina/NO, NO/GMPc, canais de K+ sensíveis ao ATP, colinérgico-muscarínico, α2-adrenérgico e gabaérgico, além de envolver a participação do glutamato e capsaicina neste efeito antinociceptivo.

Os produtos cosméticos e farmacêuticos contendo componentes de origem natural têm aumentado significativamente nos últimos anos. Com o objetivo de utilizar esses componentes como conservante em formulações cosméticas, determinou-se a atividade antimicrobiana do extrato bruto de folhas de Rubus rosaefolius Sm. e suas frações. Aquele de melhor desempenho foi avaliado quanto à toxicidade in vitro e seu comportamento em formulações cosméticas (creme, gel e xampu), relacionado à estabilidade, eficácia do sistema conservante e compatibilidade epidérmica. A atividade antimicrobiana foi determinada pelo método de microdiluição e a concentração de 0,2% (p/v) do extrato bruto apresentou melhor desempenho. O teste de toxicidade do extrato bruto foi realizado por meio do método colorimétrico Cell Titer 96®, MTS, em cultura de queratinócitos humanos, constatando-se índice de citotoxicidade (IC50) de 1,0 mg/mL. As formulações cosméticas contendo o extrato foram analisadas quanto à estabilidade e as de melhor desempenho foram submetidas ao teste de eficácia do sistema conservante, de acordo com os procedimentos descritos na CTFA. O sistema conservante mostrou-se efetivo frente à Escherichia coli IAL 2393 (ATCC 10536), a Pseudomonas aeruginosa IAL 1874 (ATCC 9027), ao Staphylococcus aureus IAL 1875 (ATCC 6548), a Burkholderia cepacia IAL 1834 ATCC (17759) e a Candida albicans IAL 1611 (ATCC 10231). Avaliouse a compatibilidade epidérmica das formulações em equivalentes dermo-epidérmicos, sistema tridimensional cultivados na superfície ar-líquido. Os resultados mostraram que houve diferenças na compatibilidade epidérmica dependendo das características dos componentes das formulações. Concluiu-se que o extrato bruto de folhas de Rubus rosaefolius Sm. a 0,2% (p/v) pode ser utilizado como candidato a conservante em formulações cosméticas, sendo estável e apresentar compatibilidade epidérmica.
Cosmetic and pharmaceuticals products contained natural compounds have increased in the last few years. To verify the use of these compounds as preservative in formulations, the antimicrobial activity from the raw extract of the Rubus rosaefolius Sm. leaves and its fractions was determined. The toxicity in vitro and the behavior in cosmetic formulations (gel, emulsion and shampoo), regarding to the stability, effectiveness of the preservative system, and epidermal compatibility were evaluated in the extract, which had the best preservative action. The antimicrobial activity was determined by the micro dilution method, and the 0.2% concentration (w/v) of the raw extract had the best performance. The toxicity of the extract was analyzed by Cell Titer 96® colorimetric; MTS method in human keratinocytes culture, and the index of cytotoxicity (IC 50) found was 1.0 mg/mL. The cosmetic formulations with the raw extract were analyzed regarding to the stability and the best formulations were submitted to preservative challenge test, according to CTFA procedures. The preservative system was effective against Escherichia coli IAL 2393 (ATCC 10536), Pseudomonas aeruginosa IAL 1874 (ATCC 9027), Staphylococcus aureus IAL 1875 (ATCC 6548), Burkholderia cepacia IAL 1834 ATCC (17759), and Candida albicans IAL 1611 (ATCC 10231). The epidermal compatibility of the formulations was verified by skin (dermo-epidermal) equivalent, three-dimensional system, cultivated in the air-liquid surface. The results showed there were differences in the epidermal compatibility depended on the ingredient of formulation. In conclusion, the raw extract from the Rubus rosaefolius Sm leaves 0.2% (w/v) can be used as preservative candidate in the cosmetic formulations analyzed due to stability and it presented epidermal compatibility.

O jambeiro (Syzygium jambos (L.) Alston) constitui uma das diversas espécies frutíferas e medicinais pertencentes à família Myrtaceae. O extrato hidroetanólico a 70% liofilizado de folhas de S. jambos apresentou atividade dose-dependente em modelo de úlcera gástrica induzida por etanol acidificado, sendo que a dose de 400 mg/kg reduziu significativamente a Área Total de Lesão (81,64%) e a Área Relativa de Lesão (65,11 %), em comparação ao grupo controle. Nesta dose, o extrato apresentou-se mais eficaz que o fármaco empregado como referência (lansoprazol 30 mg/kg). No modelo de indução de úlcera gástrica por ácido acético, o extrato (400 mg/kg) não apresentou resultados significativos na cura das lesões. A atividade antioxidante do mesmo extrato e de quatro frações foi avaliada através da medida da capacidade seqüestrante de radicais 1,1-difenil-2-picrilidrazila. O extrato hidroetanólico a 70% liofilizado apresentou CE50 de 5,36 ± 0,06 µg/mL, valor comparável ao do Trolox (CE50 =4,98 ± 0,04 µg/mL) , substância antioxidante de referência. As frações clorofórmica, acetato de etila, etanólica e hidroetanólica a 50% apresentaram CEso de, respectivamente, 64,06 ± 0,68 µg/mL, 19,02 ± 0,22 µg/mL, 6,89 ± 0,12 µg/mL e 8,47 ± 0,05 µg/mL. Da fração clorofórmica do extrato foi isolado o triterpeno ácido ursólico. Na avaliação da toxicidade subcrônica através da administração oral a ratos Wistar, durante 30 dias, de três diferentes doses do extrato (400, 1000 e 2500 mg/kg), não foram observados sinais clínicos de toxicidade. As análises macroscópica e microscópica dos órgãos não mostraram alterações dignas de nota para nenhuma das doses empregadas. Não houve diferenças estatisticamente significativas nos resultados das análises bioquímicas do sangue dos animais. Os resultados revelam o potencial do extrato no tratamento de úlceras gástricas, sendo necessários estudos mais aprofundados do mecanismo de ação desta atividade, bem como de toxicidade crônica.
Syzygium jambos (L.) Alston is one of the species of Myrtaceae with medicinal properties. The dried 70% hydroethanolic extract of the leaves showed a doseresponse effect in ethanol/HCI-induced ulcers, significantly decreasing the total lesion area (81,64%) and relative lesion area (65,11 %) compared to control group. At this dose the extract was more effective than lansoprazol (30 mg/kg), used as reference drug. The same extract at 400 mg/kg was not effective on healing acetic acid-induced ulcers. Antioxidant activity of S. jambos extract and four fractions was measured using 1,1-diphenyl-2-picryl-hydrazyl radical scavenging ability. The EC50 value for the extract was 5,36 ± 0,06 µg/mL, while Trolox, antioxidant substance of reference, showed EC50 of 4,98 ± 0,04 µg/mL. The tested fractions (chloroform, ethyl acetate, ethanol and 50% ethanol) showed EC50 of 64,06 ± 0,68 µg/mL, 19,02 ± 0,22 µg/mL, 6,89 ± 0,12 µg/mL e 8,47 ± 0,05 µg/mL, respectively. Ursolic acid was identified in the chloroformic fraction of the extract. Subchronic toxicity studies were performed by oral administration of the leaf extract to rats, during 30 days, at three different doses (400, 1000 and 2500 mg/kg). The extract did not show any clinicai sign of toxicity. Macroscopic and microscopic analysis of the organs demonstrated no alterations for the three doses tested. There were no statistically differences between results of biochemical analysis of blood. These results show the potential of the extract in treatment of gastric ulcer, although more studies of mechanism of action and chronic toxicity are necessary.

Information about the medicinal uses of forty-two plant species was collected from traditional healers and knowledgeable villagers from a variety of different ethnic groups in Nepal. Illnesses for which these plants are used are those perceived in western style medicine to be caused by bacterial, fungal or viral pathogens. Methanol extracts of the species were screened for activity against a variety of bacteria, fungi and viruses, under various light conditions to test for photosensitizers. Thirty-seven extracts showed activity against bacteria and thirty-five showed activity against fungi. Only eight were active against Gram-negative bacteria. The exposure to UV-A light had a considerable effect on the activities of some extracts, with eight extracts being active only when exposed to light. The antibacterial and antifungal effects of fifteen extracts were enhanced upon exposure to light. Fifteen extracts showed 100% inactivation of at least one virus, and fifteen showed partial activity. Eight extracts were active only when exposed to light, and the antiviral effect of eight extracts was enhanced upon exposure to light. A species showing antibacterial activity, Centipeda minima (Asteraceae), and one showing antiviral activity, Carissa carandas (Apocynaceae) were the focus of bioactivity guided fractionation. Centipeda minima was found to contain three sesquiterpene lactones, identified as 6-O-methylacrylylplenolin, 6-O-isobutyroylplenolin, and 6-O-angeloylplenolin. 6-O-Methylacrylylplenolin had not been previously isolated from C. minima. All three of these sesquiterpene lactones had activity against Bacillus subtilis and Staphylococcus aureus. A fraction from the methanol extract of Carissa carandas was quite active against herpes simplex virus. This fraction was found to contain a derivative of 3,4,5-trimethoxycinnamic acid.

Among all the major infectious human diseases, gastro-intestinal infections caused by microbial pathogens are a major cause of morbidity and infant death in developing countries, largely due to inadequate sewage disposal and contaminated water. Traditional health practitioners in South Africa play a crucial role in providing health care to the majority of the population. Many plants are locally used by South African traditional healers to treat microbial infections related to gastro-intestinal tracts. Ethnopharmacological and ethnobotanical studies using traditional knowledge as a selection strategy has given priority to certain plants for isolation and identification of plant novel bioactive compounds. Pharmacological and phytochemical studies of the investigated twelve medicinal plant species (from 10 families) extensively used as antimicrobials against gastro-intestinal infections was necessary to validate the use of the plants. Furthermore, to provide sufficient preliminary information for the isolation and identification of active compounds that are present in the investigated plants. Plant parts were sequentially extracted using petroleum ether (PE), dichloromethane (DCM) and 70% ethanol (EtOH). Cold water and boiled (decoction) extracts of the plant materials were prepared non- sequentially. Among the extracts, EtOH yielded the highest amount of plant substances. A total number of 85 extracts were evaluated for antibacterial activity, 80 for antifungal activity, 64 for anti-inflammatory activity, and 27 biologically active extracts were tested for genotoxicity. The microdilution method was used to determine the minimum inhibitory concentration values in the antibacterial assay against two Gram-negative bacteria (Escherichia coli ATCC 11775 and Klebsiella pneumoniae ATCC 13883) and two Gram-positive bacteria (Bacillus subtilis ATCC 6051 and Staphylococcus aureus ATCC 12600). A modified microdilution method was used to determine the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values in the antifungal assay against Candida albicans. Cyclooxygenase assay was used to evaluate the anti-inflammatory activity of the extracts against cyclooxygenase-1 and -2 (COX-1 and COX-2) enzymes. The plant extracts were screened first at a concentration of 250 ƒÊg/ml per test sample, and then further screened at concentrations of 125 and 62.5 ƒÊg/ml for extracts that inhibited the COX-2 enzyme. The Ames test was used to test for genotoxicity in extracts that showed interesting pharmacological activities using Salmonella typhimurium strain TA98. Among the screened extracts, 25 extracts showed good antibacterial activity with MIC values . 1.0 mg/ml. Dichloromethane extracts exhibited the greatest antibacterial activity, and Gram-positive bacteria were most susceptible. The best antibacterial activity was exhibited by Becium obovatum leaf EtOH extracts with an MIC value of 0.074 mg/ml. A broad spectrum antibacterial activity was observed by leaf extracts of Cucumis hirsutus (PE), Haworthia limifolia (PE), Protea simplex (PE and DCM) and Dissotis princeps (EtOH) against both Gram-negative and Gram-positive bacteria. No interesting antibacterial activity was exhibited by water extracts with the exception of Dissotis princeps water extract with a good antibacterial activity against Gram-positive and Gram-negative bacteria. In the antifungal assay, 6 extracts showed interesting antifungal activity. Protea simplex leaf PE extract showed the best fungicidal activity with an MFC value of 0.014 mg/ml. The best overall antifungal activity was observed in plant EtOH extracts. Some extracts from Agapanthus campanulatus (leaves and roots), Dissotis princeps (leaves), Gladiolus dalenii (corms) and Protea simplex (leaves) showed good activity against Candida albicans. Twenty one extracts inhibited the COX-1 enzyme, while fifteen extracts inhibited the COX-2 enzyme at the lowest screening concentration of 62.5 ƒÊg/ml. The highest COX-1 inhibition at a concentration of 62.5 ƒÊg/ml was exhibited by Diospyros lycioides leaf PE extract (89.1%) while Agapanthus campanulatus root DCM extract showed the highest COX-2 inhibitory activity (83.7%) at the same concentration. In the Ames test, no genotoxicity was observed in any of the extracts, however more tests need to be done to confirm these results. Thin layer chromatograms of the organic solvent plant extracts were developed. The fingerprints of the plant extracts showed colours of bands at different Rf values when viewed under UV254 and UV366 suggesting that the investigated plant species contained different compounds in the extracts. In the quest to understand the source of the plants pharmacological activities, total phenolic compounds including condensed tannins, gallotannins and flavonoids were quantitatively investigated in terms of their amounts in the aqueous methanol extracts of the plants materials using spectrophotometric methods. Alkaloids and saponins were qualitatively determined. The amounts of total phenolics were determined by the Folin Ciocalteu assay, condensed tannins were determined by the butanol-HCl assay, while rhodanine and vanillin assays were used to determine the amounts of gallotannins and flavonoids respectively. Dragendorff reagent was used to detect alkaloids in the plant extracts on thin layer chromatographic plates, while the froth test was employed to detect saponins. Secondary metabolites varied with plant parts and species with Cyperus textilis (leaf) having the highest amounts of total phenolics, condensed tannins and flavonoids. The highest amount of gallotannins was detected in Protea simplex leaf extracts. All the investigated plant materials with the exception of Haworthia limifolia leaf, Protea simplex leaf, Antidesma venosum leaf and Dissotis princeps leaf tested positively to saponins. Alkaloids were detected in Haworthia limifolia leaf (PE and EtOH), Cucumis hirsutus leaf (EtOH), Becium obovatum root (DCM), Protea simplex root and bark (EtOH), Agapanthus campanulatus root (DCM) and leaf (EtOH), Cyperus textilis root (DCM), Vernonia natalensis leaf (PE), Antidesma venosum leaf (PE), Diospyros lycioides leaf (PE) and Dissotis princeps leaf (DCM) extracts. The results obtained from the investigation of the pharmacology and phytochemistry of the plant species used to treat microbial infections related to gastro-intestinal tracts, provide sufficient preliminary information to validate the use of some of the plants in traditional medicine. The information provided might be considered sufficient for further studies aimed at isolating and identifying the active compounds in the plant species, and evaluating possible synergism amongst the isolated compounds.
Thesis (M.Sc)-University of KwaZulu-Natal, Pietermaritzburg, 2009.

Traditional medicine in South Africa is part of the culture of the people and has been in existence for a long-time. Although animal components form part of the ingredients used, plant material constitutes the major component. South Africa is endowed with vast resources of medicinal and aromatic plants which have been employed for treatment against various diseases for decades. A large number of South Africans still depend on traditional medicine for their healthcare needs due to its affordability, accessibility and cultural importance. Helminth infections are among the variety of diseases treated by traditional healers. These infections are regarded as neglected tropical diseases (NTDs) due to their high prevalence among the economically disadvantaged living in rural areas in different regions of the world.
Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2009.

Halenia elliptica D. Don belongs to Gentianaceae family. It is often used as part of a traditional Tibetan medicine to treat hepatitis. In the present investigation, six major xanthone components were isolated and identified from Halenia elliptica. An HPLC/DAD/APCI/MS method was developed and validated for the quantitative analysis of these xanthones, including 1-hydroxy-2,3,5-trimethoxy-xanthone (HM-1), 1-hydroxy-2,3,4,7-tetramethoxy- xanthone (HM-2), 1-hydroxy-2,3,4,5-tetramethoxy-xanthone (HM-3), 1,7- dihydroxy-2,3,4,5-tetramethoxy-xanthone (HM-4), 1,5-dihydroxy-2,3-dimethoxy-xanthone (HM-5) and 1-0-[beta-D-xylopyranosyl-(1-6)-beta-D-glucopyranosyl]-2,3,5-trimethoxy-xanthone (HM-2-10). All the xanthones aglycons caused vasodilation in the coronary artery pre-contracted with 1 muM 5-HT, but the xanthone glycoside had no effect. HM-1 was one of the most abundant xanthones with the most potent vasorelaxant activity.
Mechanisms of the vasorelaxant effect of HM-1 were investigated. HM-1 showed a potent vasorelaxant activity on rat coronary artery involved both an endothelium-dependent mechanism involving NO and an endothelium-independent mechanism by inhibiting Ca2+ influx through L-type voltage-operated Ca2+ channels.
Taken together, in spite of the pharmacokinetics results showed that HM-1 was rapidly and widely distributed to tissues after intravenous administration in rats, with conjugation to being the major metabolic pathway in vivo, both HM-1 and its active metabolite (HM-5) show that they are important pharmacological agents with potentially useful therapeutic indications.
The metabolism and pharmacokinetics of HM-1 displayed biphasic elimination kinetics, with an elimination half-life of 60.4 +/- 4.2 min. Four other Phase I metabolites were isolated and identified as demethylated products in vitro. HM-1 was metabolised to HM-5 in the liver. Biliary excretion studies showed that both HM-1 and the metabolite (HM-5) underwent extensive phase II conjugation to form glucuronides and sulfates. Tissue distribution studies showed that HM-1 was widely distributed to different organs. Collection of urine and faeces over 24 h showed that 10.88% of dose was excreted from urine and 1.91% of dose via faeces.
With HM-5 being one of the major in vivo metabolites of HM-1, the effect of HM-5 has been studied on rat coronary artery and compared to HM-1. HM-5-mediated vasorelaxant effect was mediated through opening of potassium channel (TEA, 4-AP) and altering intracellular calcium by partial inhibition of Ca2+ influx through L-type voltage-operated Ca 2+ channels and intracellular Ca2+ stores.
"September 2007."
Source: Dissertation Abstracts International, Volume: 69-08, Section: B, page: 4699.
Thesis (Ph.D.)--Chinese University of Hong Kong, 2007.
Includes bibliographical references (p. 195-218).
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Abstracts in English and Chinese.
School code: 1307.

Petroleum ether, dichloromethane, and 80% ethanol extracts of 15 plant species collected in Nigeria were screened for in vitro antibacterial, anti-inflammatory and antimalarial activities. Antibacterial activity was tested using the agar diffusion method, while the minimum inhibitory concentrations (MIC) of the active extracts were determined using the microtitre serial dilution method. Most antibacterial activity detected was against Gram-positive bacteria with Staphylococcus aureus being the most susceptible. The highest activity was found in petroleum ether and dichloromethane leaf extracts of Mallotus oppositifolius; petroleum ether, dichloromethane and ethanolic root extracts of Newbouldia laevis; and ethanolic root extracts of Morinda lucida and Canthium subcordatum. Against the Gram-negative bacterium Escherichia coli, the highest activity was found in dichloromethane leaf extracts of Newbouldia laevis, ethanolic root extracts of Phyllanthus amarus, Mallotus oppositifolius, and Canthium subcordatum. A total of 60 plant extracts were screened for antiplasmodial activity. A chloroquine sensitive strain of Plasmodium falciparum (D10) was used. In the assay, the parasite lactate dehydrogenase (pLDH) activity was used to measure parasite viability. About 11 extracts showed promising activity with an IC₅₀ ranging from 2.5 to 13.4 µg/ml. The petroleum ether leaf extract of Hyptis suaveolens had the highest activity (IC₅₀ = 2.5 µg/ml). The cyclooxygenase (COX-1 and COX-2) assays were used to test for anti-inflammatory activity. All the plant species, with the exception of Hedranthera barteri and Picralima nitida showed anti-inflammatory activity. Apart for a few ethanolic extracts, all the activities were recorded with petroleum ether and dichloromethane extracts. Employing bioassay-guided activity fractionation, an antibacterial anthraquinone identified as emodin was isolated from ethanolic root extract of Senna occidentalis. Although this compound had been isolated from other sources, this was the first report of isolation from Senna occidentalis. Using a similar approach a novel antimalarial diterpenoid was isolated from the petroleum ether leaves extract of Hyptis suaveolens. It had IC₅₀ of 0.1 µg/ml. This new compound is worthy of further investigation and may act as an important lead compound for future antimalarial drugs.
Thesis (Ph.D.)-University of KwaZulu-Natal, 2005.

Background and Aims of Study. Scilla nervosa is a member of the Hyacinthaceae plant family that has been naturalised in the grasslands of Southern Africa. The bulbs are traditionally used to treat a variety of ailments. For example, the Zulu people use aqueous decoctions of the bulbs as analgesics in the treatment of rheumatic fever, crushed bulbs are used by the Sotho people as laxatives and the Tswana people use cooked bulbs to treat infertility in women as well as cold aqueous extracts to treat infections. It was recently demonstrated in our laboratory that extracts prepared from the bulbs possess potent anti-inflammatory properties, and this may therefore provide a rationale for the traditional use of the plant as an analgesic. Several studies have demonstrated that the bulbs contain homoisoflavanones and stilbenoids that could be responsible for their therapeutic effects. Although the plant has diverse medicinal applications, and despite it being recognised as a poisonous species particularly in livestock, little is known about its toxicity in human liver cells. The objectives of this study were therefore to investigate the potential toxicity of the bulbs on the liver, a major detoxifying organ. A human liver cell line was treated with an aqueous extract of the bulbs to investigate (1) cell viability, (2) potential mechanisms of cytotoxicity, (3) DNA integrity, and (4) changes in the cytochrome P450 enzyme activity. Materials and Methods. This study was conducted on the cultured HepG2 human hepatocellular carcinoma cell line, a model system to investigate the cytotoxicity of xenobiotics. The viability of cultured HepG2 liver cells in the presence of varying concentrations of an aqueous extract of the bulbs was determined after 24 hours treatment, and the concentration that reduced viability to 50% (IC50) was derived. Potential mechanisms of cytotoxicity at the IC50 were investigated. These included changes in metabolic activity (intracellular adenosine triphosphate (ATP) quantification), apoptosis induction (phosphatidylserine (PS) externalisation, caspase-8 and -9 induction and changes in mitochondrial membrane potential), and oxidative damage via free radical formation (lipid peroxidation). Genotoxicity was investigated by determining changes in DNA integrity (DNA fragmentation). The ability of the extract to stimulate or {¹ Du Toit, K., Kweyama, A., Bodenstein, J. 2011. Anti-inflammatory and antimicrobial profiles of Scilla nervosa (Burch.) Jessop (Hyacinthaceae). South African Journal of Science, 107:96-100.} inhibit enzymes commonly involved with drug metabolism was investigated by determining cytochrome P450 3A4 (CYP3A4) activity. Results Cell-viability decreased in a concentration-dependent manner and the IC50 was determined as 0.03 mg/ml. Treating the cells at the IC50 resulted in (1) a 1.2-fold increase in intracellular ATP levels, (2) no significant change in PS externalisation, (3) a 1.3-fold increase in caspase-8 activity, (4) a 1.1-fold decrease in caspase-9 activity, (5) no significant change in mitochondrial membrane potential, (6) a 1.9-fold increase in lipid peroxidation, (7) evidence for genotoxicity as demonstrated by DNA fragmentation, and (8) no evidence of changes in CYP3A4 activity. Conclusion. Results suggest that HepG2 liver cells are sensitive to an aqueous extract of the bulbs of S. nervosa. The extract has the potential to (1) induce apoptosis, (2) increase oxidative stress and (3) cause genotoxicity in vitro. peroxidation, genotoxicity
Thesis (M.Sc.)-University of KwaZulu-Natal, Westville, 2011.

The crisis of newly emerging diseases and the resistance of many pathogens to currently used drugs, coupled with the adverse side-effects of many of these drugs have necessitated the continuous search for new drugs that are potent and efficacious with minimal or no adverse side-effects. The plant kingdom is known to contain many novel biologically active compounds, many of which could potentially have a higher medicinal value when compared to some of the current medications. Indeed, the use of plants in traditional medicine, especially in African communities, is gaining more importance due to their affordability and accessibility as well as their effectiveness. Exponential population growth rates in many developing countries has resulted in heavy exploitation of our plant resources for their medicinal values. In addition, plant habitat destruction arising from human developmental activities has contributed to the fragmentation or loss of many plant populations. Owing to these factors, many plant species with horticultural and/or medicinal potential have become either extinct or are threatened with extinction. These threatened species cut across different taxonomic categories including shrubs, trees and succulents. Without the application of effective conservation strategies, the medicinal and/or horticultural potential of such threatened species may be totally lost with time. The extinction of such species could lead to the loss of potential therapeutic compounds and/or genes capable of being exploited in the biosynthesis of new potent pharmaceutical compounds.
Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2009.

Xing Hongtao.
"February 2003."
Thesis (Ph.D.)--Chinese University of Hong Kong, 2003.
Includes bibliographical references (p. 149-176).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Electronic reproduction. Ann Arbor, MI : ProQuest Information and Learning Company, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Mode of access: World Wide Web.
Abstracts in English and Chinese.

by Lam Tin Lun.
"July, 2003."
Thesis (Ph.D.)--Chinese University of Hong Kong, 2003.
Includes bibliographical references (p. 308-327).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Mode of access: World Wide Web.
Abstracts in English and Chinese.

Schistosomiasis is an important public health issue for rural communities located near,or around slow moving water bodies in the tropical and subtropical areas. Successful control of the disease involves multifaceted approaches, which include snail control, environmental sanitation, health education and chemotherapy. Although snail control might be an effective method of controlling schistosomiasis, there has been a general lack of control initiatives, largely due to the cost of available molluscicides. Plants offer a wide array of compounds which, on extraction, may show molluscicidal activity. If molluscicidal compounds that occur in indigenous plants can be extracted using local labour and simple technology, then there should be culturally acceptable and inexpensive molluscicides. The aim of this study was, therefore, to screen some Zulu medicinal plants for molluscicidal activity. We have also attempted to isolate the active chemical compounds from such plants. Aqueous and methanolic crude extracts of ten (10) Zulu medicinal plants, used for different medicinal and domestic purposes, were screened for molluscicidal activity on Biomphalaria pfeifferi and Bulinus africanas snails reared in the laboratory during the time of bioassay. Bayluscide® (niclosamide) was used as a positive control for comparison, while de-chlorinated tap water was used as the negative control. Six of the plants were not active against the snails. Extracts from four of the plants demonstrated weak to moderate molluscicidal activities. These plants are: (i) Sclerocarya birrea stembark, (ii) Psidium guajava (hybrid) leaves, (iii) Leonotis leonurus aerial parts and (iv) Ekerbegia capensis stem-bark. The LC50 values of the plant extracts were 78 ppm, 100 ppm, 398 ppm and 600 ppm respectively. Of the 4 plants that showed molluscicidal activity, S. birrea aqueous and methanol extracts were the most active against the snails, with LC50 values of 82 ppm and 78 ppm respectively. For the other plant extracts, only the methanolic extracts showed activity. Brine shrimp toxicity assay was performed with all the active extracts. Psidium guajava showed 10% survival of the shrimps at 1000 ppm, whereas no survival was observed for the other plant extracts at this concentration (1000 ppm). The results obtained in this study indicate that further studies have to be conducted, especially with S. birrea extracts, whose both aqueous and methanolic extracts showed significant activity against the snails.
Thesis (M.Med.Sc.)-University of Durban-Westville, 2002.

This research study undertook to investigate and evaluate for efficacy and safety, the herbal remedies used for treating Diabetes mellitus in northern KwaZulu-Natal. In addition, it sought to gain knowledge and better understanding of traditional healing systems and the medicinal use of the natural flora. During the process of assimilating the desired information, the epidemiological and socio-economic factors which determine the form of medicine chosen by rural people in the region, were quantified. Both aspects of explanatory studies i.e. experimental and observational were used. Firstly, to evaluate the safety of the two herbal remedies, laboratory animals were given an oral dose of the herbal medicine and observed for a period of 14 days. Efficacy was assessed by treating Streptozotocin-induced diabetic rats with the herbal remedies and comparing their effect on blood glucose with that of a conventional sulphonylurea. The second part of the study was observational and it involved monitoring human subjects (patients) for twelve months, who were already taking the herbal preparations (n=56) and comparing their prognoses with that of a group taking conventional medicine (n=97). A third group using both types of medicine (n=42) was included as control measure for a possible confounding factor. Main outcome measures; Both subjective and objective measures of the perceived health of the diabetic patients were measured, as well as the determinants of using traditional medicine versus conventional medicine. The battery of toxicity tests which utilises behavioural and functional observations of the laboratory animals, yielded no signs of toxicity or abnormal behaviour. The histopathological examination results of the sample organs from the treated rats also revealed no signs of abnormality that could be attributed to the herbal remedies tested. There was no sex variation recorded in the response. The first HP tested (HP-1) demonstrated minimal hypoglycaemic effect whereas HP-2 significantly lowered the blood glucose of the streptozotocin-induced diabetic rats by an average of 59%. This was comparable to the conventional medicine (Glibenclamide) used in the experiment. After 12 months of follow-up, 93 % of traditional medicine users (n=56) were convinced that their blood sugar was controlled because of the traditional remedy they were using. The proportion of diabetic cases who used conventional medicine were no better off than those who used traditional medicine or vice versa. Health status and the financial situation (income) of the respondents greatly influenced their choice for diabetic treatment. The herbal remedies that were investigated were non-toxic and safe for use and internal consumption. One preparation demonstrated a significant hypoglycaemic effect, which was comparable to the conventional allopathic medicine used in treating Diabetes mellitus. This study should serve as a springboard to encourage more pharmacological evaluation of herbal medicines.
Thesis (Ph.D.)-University of Durban-Westville, 1998.

Plants have formed the foundation of traditional medicine systems throughout the world for thousands of years and continue to provide mankind with new remedies for various ailments. A large portion of the black South African population still depends on medicinal plants as primary health care due to its affordability, accessibility and cultural importance. These medicinal plants need to be investigated since new lead compounds are often found in nature. Homoisoflavanones isolated from South African and Indian plants were found to exhibit anti inflammatory activities although the mechanism of action has not yet been determined. A few reports on the anti fungal activities of these compounds were also found. Four new and three known homoisoflavanones of the 3-benzylidene-4-chromanone type were synthesized and tested for anti-inflammatory and antifungal activities. Two novel intermediates were also synthesised. Enantiomers of a homoisoflavanone of the 3-benzyl-4- chromanone types were also synthesized from the corresponding 3,5-dimethoxy phenol via 4- chromanone in six steps. This is the first report of the synthesis of an enantiomerically pure homoisoflavanone compound together with its opposite isomer. The enantiomers and racemate were tested for anti-inflammatory activity. All the synthesized homoisoflavanones were screened for cytotoxicity. The structures of these homoisoflavanones were elucidated by NMR spectroscopy along with HRMS data. The crystal structure of a homoisoflavanone with anti-inflammatory and antifungal activity is reported. The anti-inflammatory activity of the homoisoflavanones was determined in an acute croton oil-induced auricular dermatitis mouse model. The antifungal activity was performed in vitro against a Candida albicans strain. Compounds were tested for cytotoxicity against a Chinese Hamster Ovarian (CHO) cell line using the 3-(4,5-dimethylthiazol-2-yl)-3,5- diphenyltetrazoliumbromide (MTT) assay. In conclusion, the synthetic homoisoflavanones showed anti-inflammatory as well as antifungal activity. Some of the compounds showed anti-inflammatory activity comparable to that of the commercially available diclofenac.
Thesis (Ph.D.)-University of KwaZulu-Natal, Westville, 2011.

Two South African medicinal plants, Strophanthus speciosus and Eucomis montana, were investigated phytochemically. From Strophanthus speciosus a cardenolide, neritaloside, was isolated, whilst Eucomis montana yielded three homoisoflavanones, 3,9- dihydroeucomin, 4'-demethyl-3,9-dihydroeucomin, and 4'-demethyl-5-0-methyl-3,9- dihydroeucomin. The structures were elucidated on the basis of spectroscopic data. The homoisoflavanones were screened for anti-inflammatory activity using a chemiluminescent luminol assay, modified for microplate usage. All of the homoisoflavanones exhibited good inhibition of chemiluminescence, with IC50 values for 3,9-dihydroeucomin, 4'-demethyl-3,9-dihydroeucomin, and 4'-demethyl-5-0-methyl-3,9- dihydroeucomin being 14mg/mL, 7mg/mL, and 13mg/mL respectively. The IC50 value of 4'-demethyl-3,9-dihydroeucomin compared favourably with the NSAID control (meloxicam), which had an IC50 of 6mg/mL. Neritaloside was not screened for biological activity as the yield of 14.4mg was insufficient for the muscle-relaxant screen for which it was intended. An assay for antioxidant/free radical scavenging activity was also performed. All the compounds had excellent antioxidant/free radical scavenging activity, with percentage inhibition of the reaction being 92%, 96%, and 94% for 3,9-dihydroeucomin, 4'-demethyl- 3,9-dihydroeucomin, and 4'-demethyl-5-0-methyl-3,9-dihydroeucomin respectively at a concentration of 10mg/mL. However, the control compounds, diclofenac and meloxicam, also exhibited strong activity, with the result that the precise mode of anti-inflammatory activity could not be unequivocally determined. The results from the biological screenings thus provided a rational scientific basis for the indigenous ethnomedicinal use of Eucomis species in the treatment of rheumatism, inflammation and pain.
Thesis (M.Sc.)-University of KwaZulu-Natal, 2006.

In this study two medicinal plant species, namely Artemisia afra Jacq. ex. Willd and Elytropappus rhinocerotis Less. (L.f) (= Dicerotamnus rhinocerotis Koekemoer), both belonging to the family Asteraceae, have been investigated and different compounds isolated and characterised. Both species are important plants used in traditional medicine in general in Africa and particularly in South Africa. A. afra, commonly called "African wormwood" is one of 400 species belonging to the genus Artemisia and it is the only one indigenous to Africa. E. rhinocerotis is one of eight Elytropappus species which are all restricted to the Cape floristic region. The aim of this study was to investigate the phytochemistry of these species. In total fifteen compounds were isolated and characterised. From the E. rhinocerotis extract, four known compounds, labdanolic acid, methyl labdanolate, 6, 7- dimethoxycoumarin and a sesquiterpene viridiflorol were isolated. These compounds were not previously reported from E. rhinocerotis. Two different chemotypes of A. afra were studied and eleven compounds were isolated. These compounds include sesquiterpenes such as taurin, artesin, maritimin, artemin, norsantolinifolide, santolinifolide A, and reynosin, a flavonoid, 5- hydroxy-7,4-dimethoxyflavone, a coumarin called scopoletin or 7-hydroxy-6- methoxycoumarin and other aromatic compounds such as p-hydroxyacetophenone, and 2,4-dihydroxy-6-methoxyacetophenone. Except for taurin, scopoletin and 5-hydroxy-7, 4- dimethoxyflavone , none of these other compounds has been reported previously from A. afra. This study has shown that A. afra contains a large number of sesquiterpenoids, mostly from the eudesmane-type. Structural elucidation of different compounds was performed using mainly NMR spectroscopy. Other methods used for identification include LC-MS and infrared spectroscopy. The major compound, labdanolic acid, is known to selectively inhibit cyclooxygenase-2, an enzyme associated with inflammation. The presence of labdanolic acid in the plant may account for its traditional use as an anti-inflammatory.
Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2009.

by Cheng Yuk Kwan, Anna.
Thesis (M.Phil.)--Chinese University of Hong Kong, 1993.
Includes bibliographical references (leaves 104-112).
Acknowledgements
Table of Contents --- p.i
Abbreviations --- p.iv
Abstract --- p.vi
List of figures --- p.ix
List of tables --- p.xi
Chapter Chapter One: --- Introduction
Chapter 1.1 --- Objective and scope of the project --- p.1
Chapter 1.2 --- Literature review of Mimosa pudica
Chapter 1.2.1 --- Morphology of Mimosa pudica --- p.3
Chapter 1.2.2 --- Chemistry of Mimosa pudica --- p.5
Chapter 1.2.3 --- Uses in traditional medicine --- p.5
Chapter 1.2.4 --- Clinical and pharmacological studies of Mimosa pudica --- p.6
Chapter 1.2.5 --- Toxicology of Mimosa pudica --- p.8
Chapter 1.2.6 --- Characteristics and toxicology of mimosine --- p.9
Chapter 1.3 --- Immunomodulation
Chapter 1.3.1 --- Overview of the immune system --- p.11
Chapter 1.3.2 --- Strategies on the study of immunomodulation of Mimosa pudica --- p.13
Chapter 1.4 --- Toxicology
Chapter 1.4.1 --- Principles of the toxicological assays
Chapter 1.4.1.1 --- LD50 --- p.17
Chapter 1.4.1.2 --- Enzyme assays --- p.18
Chapter 1.4.1.3 --- Subacute toxicity test --- p.24
Chapter 1.4.1.4 --- Reproductive toxicity test --- p.25
Chapter Chapter Two: --- Materials and methods
Chapter 2.1 --- Materials
Chapter 2.1.1 --- Mimosa pudica --- p.27
Chapter 2.1.2 --- Animals --- p.27
Chapter 2.1.3 --- Chemicals --- p.28
Chapter 2.2 --- Methods
Chapter 2.2.1 --- Extraction of Mimosa pudica --- p.32
Chapter 2.2.2 --- Assays for the immunomodulatory effects of Mimosa pudica
Chapter 2.2.2.1 --- Cell preparation
Chapter a) --- Splenocytes --- p.35
Chapter b) --- Thymocytes --- p.35
Chapter c) --- Macrophages --- p.36
Chapter 2.2.2.2 --- Splenocyte proliferation --- p.37
Chapter 2.2.2.3 --- Thymocyte proliferation --- p.38
Chapter 2.2.2.4 --- Phagocytic activity of macrophages --- p.39
Chapter 2.2.2.5 --- Release of IL-1 by macrophages --- p.40
Chapter 2.2.2.6 --- Plaque forming cells --- p.41
Chapter 2.2.2.7 --- Restoration on splenocyte blastogenesis of old mice --- p.42
Chapter 2.2.3 --- Assays for the toxicity of Mimosa pudica
Chapter 2.2.3.1 --- LD50 --- p.43
Chapter 2.2.3.2 --- Enzyme assays --- p.43
Chapter 2.2.3.3 --- Subacute toxicity --- p.43
Chapter 2.2.3.4 --- Reproductive toxicity --- p.44
Chapter 2.2.4 --- Statistical analysis --- p.44
Chapter Chapter Three: --- Results
Chapter 3.1 --- Immunomodulatory effects of Mimosa pudica
Chapter 3.1.1 --- In vitro study on the lymphocyte proliferation
Chapter 3.1.1.1 --- Splenocyte proliferation --- p.45
Chapter 3.1.1.2 --- Thymocyte proliferation --- p.50
Chapter 3.1.2 --- In vivo study on the lymphocyte proliferation --- p.53
Chapter 3.1.3 --- Phagocytic activity of macrophages --- p.58
Chapter 3.1.4 --- Release of IL-1 by macrophages --- p.64
Chapter 3.1.5 --- Plaque forming cells --- p.67
Chapter 3.1.6 --- Restoration on splenocyte blastogenesis of old mice --- p.69
Chapter 3.2 --- Toxicity of Mimosa pudica
Chapter 3.2.1 --- LD50 --- p.72
Chapter 3.2.2 --- Enzyme assays --- p.75
Chapter 3.2.3 --- Subacute toxicity --- p.80
Chapter 3.2.4 --- Reproductive toxicity --- p.85
Chapter Chapter Four: --- General discussion on the immunomodulatory effects and toxicity of Mimosa pudica
Chapter 4.1 --- Immunomodulatory effects of Mimosa pudica --- p.88
Chapter 4.2 --- Toxicity of Mimosa pudica --- p.95
Chapter Chapter Five: --- Concluding remarks --- p.99
References --- p.104
Appendix --- p.113

Orientação: Maria Lídia Palma
A Achillea millefolium L. é uma planta da família Asteraceae que está distribuída em várias partes do mundo, especialmente na Europa, Ásia e América. A prática medicinal do uso dos constituintes do milefólio é historiada desde a Mesopotâmia, e ao longo dos anos o intenso estudo que tem sido objeto é devido à sua composição e consequentemente à sua extensa atividade em diversas patologias/sintomatologia. Os constituintes fitoquímicos do milefólio são o óleo essencial, os alcaloides, os flavonoides, as lactonas sesquiterpénicas, os triterpenos e esteróis, os taninos, as cumarinas, o ácido salicílico e ácido ascórbico, os poliacetilenos, os carotenoides, proteínas, resinas e carboidratos. Para a extração do óleo essencial a técnica utilizada é a steam distillation e posteriormente a análise é feita por cromatografia gasosa acoplada à espectrometria de massas (CG-EM). O rácio dos componentes e a sua atividade estão relacionados com a geografia, estação do ano, parte da planta e o número de cromossomas. Neste estudo monográfico foi feita uma revisão bibliográfica das atividades in vitro e in vivo. A atividade antimicrobiana foi demonstrada, in vitro, com mais sensibilidade para as bactérias gram-negativas (Escherichia coli, Pseudomonas aeruginosa, Proteus mirabilis) do que gram-positivas (Staphylococcus aureus, Streptococcus pneumoniae). Os componentes responsáveis por esta atividade são em maiores quantidades as lactonas sesquiterpénicas, os terpenos e em pequenas percentagens a cânfora, o eucaliptol e o borneol. O flavonoide artemetina, presente na planta, foi isolado do extrato de diclorometano e administrado em ratos Wistar por via oral e intravenosa, em ambas as vias observou-se uma redução da resposta hipertensiva à angiotensina I com, um mecanismo muito semelhante ao de um Inibidor da Enzima de Conversão da Angiotensina (IECA). Num estudo in vivo onde foi feita uma comparação entre o extrato da planta e o óxido de zinco, foi demonstrado que os mono/sesquiterpenos da planta inibem o metabolismo do ácido araquidónico evitando a lesão tecidular. Os taninos obtidos do milefólio impermeabilizam, em pequenas doses, a mucosa do intestino prevenindo as diarreias. A apigenina e a quercetina, flavonoides presentes no milefólio, diminuíram as contrações do íleo.
Achillea millefolium L. is a plant of the family Asteraceae that is distributed in several parts of the world, especially in Europe, Asia and America. The medicinal practice of the use of yarrow constituents has been historied since Mesopotamia, and over the years the intense study that has been object is due to its composition and consequently to its extensive activity in diverse pathologies / symptomatology. The phytochemical constituents of yarrow are essential oil, alkaloids, flavonoids, sesquiterpene lactones, triterpenes and sterols, tannins, coumarins, salicylic acid and ascorbic acid, polyacetylenes, carotenoids, proteins, resins and carbohydrates. For the extraction of the essential oil the technique used is steam distillation and later the analysis is done by gas chromatography coupled to mass spectrometry (GC-MS). The ratio of the components and their activity are related to the geography, season of the year, part of the plant and the number of chromosomes. In this monographic study a bibliographical review of in vitro and in vivo activities was done. The antimicrobial activity was demonstrated, in vitro, with more sensitivity for gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa, Proteus mirabilis) than gram-positive (Staphylococcus aureus, Streptococcus pneumoniae). The components responsible for this activity are in greater quantities the sesquiterpene lactones, the terpenes and in smallpercentages camphor, eucalyptol and borneol. Flavonoid artemetin, present in the plant, was isolated from dichloromethane extract and administered to oral and intravenous Wistar rats. Both flavonoids reduced the hypertensive response to angiotensin I with a mechanism very similar to that of an Angiotensin Converting Enzyme Inhibitor (ACEI). In an in vivo study comparing plant extracts with zinc oxide, plant monkeys / sesquiterpenes have been shown to inhibit arachidonic acid metabolism by preventing tissue damage. The tannins obtained from the milfoil seal the intestinal mucosa in small doses, preventing diarrhea. Apigenin and quercetin, flavonoids present in yarrow, decreased contractions of the ileum.

The present study investigated pharmacological properties of Capsicum frutescens-derived capsaicin, including its analgesic, anti-inflammatory and coagulatory properties. The effects of capsaicin on gastrointestinal and myocardial muscles, as well as on myocardial ischaemic-reperfusion, were also investigated. Capsaicin pre-treatment in neonatal rats has been found to abolish the development of thermal hyperalgesia produced in a model of neuropathic pain in rats (Toth-Kasa et al., 1986). In addition, capsaicin sensitivity has been found to be dependent on continued presence of nerve growth factor (NGF), whose concentration increases in inflamed tissues (Bevan and Winter, 1995). By stimulating the release of excitatory amino acids (EAA); such as glutamate and neuropeptides [(CGRP, neurokinin A (NKA) and Substance P (SP)] from both the peripheral and central terminals of sensory neurones by two mechanisms (Kroll et al., 1990; Del Bianco et al., 1991; Lou et al., 1992; 1994; Woolf et al., 1994); capsaicin has been shown to produce a longer-term inhibitory effect. This is one likely mechanism for capsaicin analgesic and anti-inflammatory actions (Bleakman et al., 1990). Within the gastro-intestinal tract, SP and NKA are involved in the physiological control of several digestive functions, such as motility, fluid and electrolyte secretion, blood flow, and tissue homeostasis (Otsuka, 1993; Holzer et al., 1997). Consistent with this finding, upsurge of SP in irritable bowel syndrome (IBD) was confirmed by Mantyh et al, (1988). Pre-treatment of rats with either capsaicin or NK-1R antagonists dramatically reduced fluid secretion, mucosal permeability, and intestinal inflammation in animal models of acute and chronic inflammation (McCafferty et al, 1994; Pothoulakis et al., 1994). Capsaicin can modulate endocrine and paracrine activities, immune responses, as well as gastro-intestinal and cardiovascular functions. Moreover, up-regulation of Substance P receptors was found to be associated with chronic inflammatory conditions (De et al., 1990). Stimulation of transient receptor potential vanilloid 1 also results in the activation of nociceptive and neurogenic inflammatory responses (Rigoni et al., 2003). vi The pharmacodynamic effects of capsaicin on the cardiovascular system remain elusive. Some actions of capsaicin on the heart were attributed to an interaction at K+ channels (Castle, 1992), or liberation of neuropeptides, most notably calcitonin-gene-related-peptide (CGRP) from the vanilloid-sensitive innervation of the heart (Franco-Cereceda et al., 1988; 1991). The possibility of a direct effect of capsaicin on the heart via a cardiac vanilloid receptor (VR), or through interaction of vanilloid receptors with purinergic receptors, and subsequent release of nitric oxide (NO), leading to vasodilatation were considered. Evidence abound in the literature that Ca2+ ions are released through 1, 4, 5 inositol phosphatase by the release of phospholipase C, or through interaction of the vanilloid receptors with cannabinoids. In an earlier study, Jaiarj et al. (1998) found that capsaicin acting on the heat-sensitive vanilloid receptors, had thrombolytic effects. Though weak evidence, Jaiarj et al. (1998) observed that individuals who consume large amounts of Capsicum have lower incidence of thromboembolism. Following ethical approval, the study reported in this thesis was conducted in phases. Identification of Capsicum frutescens (facilitated by a botanist in the Department of Botany, Westville campus of the University of KwaZulu Natal). Chromatographic extraction of capsaicin from Capsicum frutescens was followed by Nuclear Magnetic Resonance (NMR) analysis of the extract. Animal studies were conducted using capsaicin extract (CFE) and/or a reference capsaicin (CPF), using „hot plate. and „acetic acid. test methods to investigate the role of capsaicin on analgesia. Fresh egg albumin-induced inflammation was used to investigate the role of capsaicin in inflammation, following pre-treatment with CFE and CPF. Concentraton-response curves of increasing concentrations of capsaicin, acetylcholine and other agonist drugs with specific antagonists on strips of chick oesophagus, guinea-pig ileum, and rabbit duodenum were constructed following investigations on gastrointestinal (GIT) smooth muscles. The effect of capsaicin on coagulation was assessed by measuring international normalized ratio (INR) of animals that were exposed to different concentrations of capsaicin (CFE and CPF). Furthermore, parallel control studies were conducted in each of these investigations using distilled water or saline as placebo-control or specific-prototype agonists. negative-control. Cardiovascular investigations included studies on the effects of capsaicin on the heart rate, inotropy, vii coronary perfusion pressure, and ischaemic-reperfusion injury, using Langendorf.s rat heart models. Collated data were triangulated by manual hand-written and PowerLab data acquisition, or computerised capture. Statistical analysis were performed by either one or two of the following: Student.s t-test, ANOVA (repeated or single–use modes), facilitated and confirmed by Graph Pad Prism, Microsoft Excel or CPSS software(s). Reproducibility and relevance to the stated objectives of the various studies were confirmed by assessing which of the Null or Alternative hypothesis is validated by the results from the test. Treatment with CFE or CPF at all doses significantly (p<0.01) increased MRT. By comparison with control, writhing responses to acetic acid were significantly reduced following pre-treatment with various doses of CFE or CPF. The results in both parallel groups of CFE and CPF in the hot plate and acetic acid tests had Pearson correlation of one (1). Compared to the diclofenac (DIC) group, the degree of inhibition of paw oedema by CFE and CPF was statistically significant (P<0.05-0.001), best in the first 4 hours of treatment. The results of the in vitro laboratory animal study indicate that relatively low concentration of CPF (20 or 40 .g) produced significant (p.0.05), concentration-related inhibitions of acetylcholine (0.1-5 .g)-induced contractions of the chick isolated oesophagus, guinea-pig isolated ileum and rabbit isolated duodenum. Biphasic effects, which were noticed at low concentrations, consisted of initial brief contractions, followed by longer-lasting relaxations and reductions of the contractile amplitudes of the muscle preparations. Percentage inhibitions of the smooth muscle contractions by CFE or CPF were concentration-dependent, ranging from 20-70% (p<0.02).
Thesis (Ph.D.)-University of KwaZulu-Natal, Westville, 2012.

Hung Sau Ling.
Thesis (M.Phil.)--Chinese University of Hong Kong, 1994.
Includes bibliographical references (leaves 131-137).
Table of Contents --- p.1
Acknowledgements --- p.V
Abbreviations --- p.VI
Aim of investigation --- p.IX
Abstract --- p.XI
Chapter Chapter 1 --- General Introduction --- p.1
Chapter 1.1 --- Cancer Chemotherapy --- p.2
Chapter 1.2 --- Plants as sources of useful drugs --- p.4
Chapter 1.3 --- Potent antitumor compounds found in Goniothalamus giganteus --- p.7
Chapter 1.4 --- Brief introduction of GONIOTRIOL --- p.8
Chapter 1.5 --- The study on the antitumor activities of the antitumor compounds --- p.9
Chapter 1.6 --- Biochemistry study of the anticancer agents --- p.10
Chapter Chapter 2 --- Materials and Methods --- p.18
Chapter 2.1 --- Materials --- p.19
Chapter 2.1.1 --- Animals --- p.19
Chapter 2.1.2 --- "Buffers, Culture Media and Chemicals" --- p.19
Chapter 2.1.3 --- Cell lines --- p.20
Chapter 2.1.4 --- Dye solutions --- p.21
Chapter 2.1.5 --- Reagents and buffers for Agarose gel --- p.21
Chapter 2.1.6 --- Synthetic goniotriol and its derivatives --- p.21
Chapter 2.2 --- Methods --- p.23
Chapter 2.2.1 --- Radioactive Precursor Incorporation Assays --- p.23
Chapter 2.2.2 --- MTT assay --- p.24
Chapter 2.2.3 --- Neutral Red assay --- p.24
Chapter 2.2.4 --- Isolation and preparation of cells --- p.25
Chapter 2.2.5 --- Assay for the solvent effect --- p.25
Chapter 2.2.6 --- Assay for the in vitro antitumor activity THC88 on different cell lines --- p.27
Chapter 2.2.7 --- Assay of the effect of THC86 on solid sarcoma Scl80 in vivo --- p.28
Chapter 2.2.8 --- Assay of the effect of THC86 on peritoneal Scl80 in vivo --- p.28
Chapter 2.2.9 --- Assay of the effect of THC89 on peritoneal EAT in vivo --- p.28
Chapter 2.2.10 --- Assay of synthetic compound (THC89 and THC87) on the mitogenic activity of spleen lymphocytes --- p.29
Chapter 2.2.11 --- Assay of synthetic compound (THC87) on the proliferation of murine bone marrow cells from compound- treated mice --- p.30
Chapter 2.2.12 --- "Assay of synthetic compounds (Ml, P51 and P1) on nonmalignant cell-line" --- p.31
Chapter 2.2.13 --- Assay of antitumor activity of synthetic compound (THC86)on PU5-1.8 --- p.31
Chapter 2.2.14 --- Assay of the cytocidal effect of THC86 --- p.32
Chapter 2.2.15 --- "Assay on the effect of THC86 on the synthesis of DNA, RNA and protein" --- p.32
Chapter 2.2.16 --- Direct DNA cleavage by THC86 --- p.33
Chapter 2.2.17 --- DNA fragmentation assay
Chapter 2.2.18 --- Assay of the effect of the synthetic compound (THC86) on different growth fraction of the cells
Chapter 2.2.19 --- Mitosis Study
Chapter 2.2.20 --- Assay for the stability of the synthetic compounds
Chapter Chapter 3 --- Structure / activity relationship of the synthetic compounds --- p.36
Chapter 3.1 --- Results --- p.37
Chapter 3.1.1 --- In vitro antitumor activity of the synthetic compounds --- p.37
Chapter 3.2 --- Discussion --- p.45
Chapter Chapter 4 --- Antitumor activities of the synthetic compounds --- p.63
Chapter 4.1 --- Results --- p.64
Chapter 4.1.1 --- Solvent effect in the screening process --- p.64
Chapter 4.1.2 --- The effect of the synthetic compound (THC88) on different cell lines --- p.69
Chapter 4.1.3 --- In vivo anti-tumor activities of the synthetic compounds --- p.71
Chapter 4.1.3a --- Effect of THC86 on solid sarcoma Sc180 in vivo --- p.71
Chapter 4.1.3b --- Effect of THC86 on peritoneal Scl80 in vivo --- p.71
Chapter 4.1.3c --- Effect of THC89 on peritoneal EAT in vivo --- p.72
Chapter 4.1.4 --- Cytotoxic effect of the tested compounds on normal cells --- p.77
Chapter 4.1.4a --- Cytotoxic effect of THC89 on normal splenocytes in vitro --- p.77
Chapter 4.1.4b --- Effect of THC87 on the proliferation of splenocytes --- p.77
Chapter 4.1.4c --- Effect of THC87 on the proliferation of murine bone marrow cells --- p.78
Chapter 4.1.4d --- Cytotoxic effect on non-malignant cell-line BALB/c 3T3/A31 --- p.78
Chapter 4.2 --- Discussion --- p.85
Chapter Chapter 5 --- The study on the antiproliferative mechanisms of the synthetic compounds --- p.88
Chapter 5.1 --- Results --- p.89
Chapter 5.1.1 --- "Effect of the synthetic compounds on Cell Growth, DNA, RNA and Protein" --- p.89
Chapter 5.1.1a --- Effect of THC86 on PU5-1.8 (macrophage-like tumor) --- p.89
Chapter 5.1.1b --- Cytocidal effect of THC86 on EAT --- p.89
Chapter 5.1.1c --- "Effect of the synthetic compounds on synthesis of DNA, RNA and protein" --- p.90
Chapter 5.1.2 --- Study of the synthetic compounds on the interactions of DNA --- p.101
Chapter 5.1.2a --- DNA cleavage assay --- p.101
Chapter 5.1.2b --- DNA fragmentation assay --- p.101
Chapter 5.1.3 --- Effect of the synthetic compounds on different growth fraction of the cells --- p.104
Chapter 5.1.4 --- Mitosis study of the synthetic compounds --- p.106
Chapter 5.1.5 --- Investigation of the stability of the synthetic compounds in culture medium --- p.112
Chapter 5.2 --- Discussion --- p.117
Chapter Chapter 6 --- General Discussion --- p.122
References --- p.131

Orientação : Amílcar Roberto
A malária é a doença parasitária mais preocupante da atualidade, sendo um grave problema de saúde pública, principalmente em África onde mata maioritariamente crianças com idade inferior a 5 anos. As estratégias farmacoterapêuticas de combate à malária, baseiam-se principalmente nos fármacos derivados da artemisinina e em fármacos quinolínicos. A artemisinina e a quinina são fármacos provenientes das espécies vegetais, Artemisia annua L. e Cinchona L., respectivamente, e são conhecidos há muito tempo pela medicina tradicional. Estas substâncias são as que têm maior importância no tratamento da malária não apresentando fármaco-resistências de larga escala, sendo que a quinina permanece eficaz, apesar de algumas perdas de susceptibilidade por parte do Plasmodium, e as artemisininas, nomeadamente os seus derivados semissintéticos artemeter, artesunato e dihidroartemisinina, apenas apresentam resistência no sudeste asiático. Desta forma é importante conhecer as características destes fármacos, de forma a perceber as vantagens que apresentam, para além da sua eficácia contra o parasita, caracterizando os seus mecanismos de acção, farmacodinâmica e cinética, posologia e toxicidade. A perda de susceptibilidade e o aparecimento de fármaco-resistências constituem um desafio, pelo que é muito importante implementar medidas para que estas não surjam em grande escala, como o controlo do vector, bons métodos de diagnóstico, prevenção, uso correto das terapêuticas e maneiras de incentivar a adopção de todas estas medidas de forma eficaz pelos países. Os princípios ativos antimaláricos da Cinchona L. e da Artemisia annua L. têm características importantes que devem ser utilizadas na procura e síntese de novos fármacos antimaláricos. A quinina permanece eficaz mesmo em estirpes de Plasmodium resistentes, e têm sido desenvolvidos trabalhos de investigação de modo a justificar esta eficácia, já que grande parte dos fármacos quinolínicos, como a cloroquina e a mefloquina, apresentam fármaco-resistências. Esta eficácia da quinina poderá estar relacionada com o seu grupo substituinte mais complexo, comparativamente ao grupo substituinte da cloroquina. As artemisininas têm sido utilizadas como modelo no desenvolvimento de novos fármacos. A sua ponte endoperóxida, considerada a grande responsável pelo efeito antimalárico, foi utilizada como base na criação de potenciais fármacos, como o arterolano.
Malaria is the most worrying parasitic disease and represents a serious public health problem, especially in Africa where it kills mostly children under five years old. The pharmacotherapeutic strategies to prevent and treat malaria are mainly based on artemisinin derivatives and quinoline drugs. Both quinine and artemisinin are obtained from vegetal species, Artemisia annua L. and Cinchona L., and their use is known a long time ago by traditional medicine. These substances have major importance in the treatment of malaria because they don’t exhibit a long scale drug-resistance, as quinine remains effective despite some loss of sensitivity and artemisinins, including artemether, dihydroartemisinin and artesunate, only show resistance in Southeast Asia. Thus it is important to know the characteristics of these two agents in order to realize their advantages, beyond its anti-parasitic effect, realizing their mechanisms of action, pharmacodynamics and kinetics, toxicity and dosage. The loss of susceptibility and the emergence of drug-resistance in these drugs represent a challenge, so it is very important to be aware of the strategies to adopt in order to not arise them on a large scale. Some of these measures include vector controlling, good methods of diagnosis, prevention, correctly using therapeutics and encouragement to implement effectively all of these measures. The antimalarial drugs of Cinchona L. and Artemisia annua L. have important features that shall be used when searching and synthetizing new antimalarial drugs. Quinine remains effective even against resistant strains of Plasmodium, and has been researched in order to justify this, since much of quinoline drugs present drug-resistance. This efficacy can be related to its more complex substituent group, compared to the chloroquine’s one. Artemisinins have been used as a model for the development of new drugs. Its endoperoxide bridge, which is considered the main feature for its antimalarical effect, has been used in the creation of new potential drugs such as arterolane.

La prévalence de l’obésité, du diabète de type 2, et du syndrome métabolique, sont à la hausse chez les Cris d’Eeyou Istchee (CEI-Nord du Québec). Ces problèmes sont aggravés par leur diète non traditionnelle, leur sédentarité, ainsi que par une résistance culturelle aux produits pharmaceutiques. Afin de développer des traitements antidiabétiques culturellement adaptés, notre équipe a effectué une enquête ethnobotanique qui a identifié 17 plantes provenant de la pharmacopée traditionnelle des CEI. À partir des études de criblage effectuées in vitro, deux plantes parmi les 17 ont attiré notre attention. Populus balsamifera L. (Salicaceae) pour ses propriétés anti-obésité et Larix laricina K. Koch (Pinaceae) pour ses propriétés antidiabétiques. P. balsamifera et son composé actif salicortin ont inhibé l’accumulation de triglycérides durant l’adipogénèse dans les adipocytes 3T3-L1. L. laricina a augmenté le transport de glucose et l’activation de l’AMPK dans les cellules musculaires C2C12, l’adipogénèse dans les 3T3-L1 et a démontré un fort potentiel découpleur (propriété anti-obésité). Les objectifs de cette thèse sont d'évaluer les potentiels anti-obésité et antidiabétique et d’élucider les mécanismes d'action de P. balsamifera, salicortin, et L. laricina chez la souris C57BL/6 rendue obèse par une diète riche en gras (HFD). Les souris ont été soumises pendant huit (étude préventive) ou seize semaines (étude traitement) à une HFD, ou à une HFD dans laquelle P. balsamifera, salicortin, ou L. laricina a été incorporé soit dès le départ (prévention), ou dans les 8 dernières des 16 semaines d'administration de HFD (traitement). iv Les résultats démontrent que P. balsamifera (dans les deux études) et salicortin (évalué dans l’étude traitement) diminuent: le poids corporel, le gras rétropéritonéal, la sévérité de la stéatose et l’accumulation de triglycérides hépatique (ERK impliqué), les niveaux de glycémie et d'insuline, et le ratio leptine/adiponectine. Dans les deux études, P. balsamifera a significativement réduit la consommation de nourriture mais cet effet coupe-faim nécessite d’être approfondi. Dans l'étude préventive, P. balsamifera a augmenté la dépense énergétique (hausse de la température à la surface de la peau et de l’activation de la protéine découplante-1; UCP-1). Les voies de signalisation activées par P. balsamifera et par salicortin (de façon plus modeste) sont impliquées dans: la production de glucose hépatique (Akt), l’expression de Glut4 dans le muscle squelettique, la captation du glucose et du métabolisme des lipides (Akt dans le tissu adipeux), la différenciation des adipocytes (ERK et PPARg), l’inflammation dans le foie (IKKαβ), et l'oxydation des acides gras dans le muscle, le foie, ou le tissu adipeux (PPARa et CPT-1). D’autre part, L. laricina a également diminué les niveaux de glycémie et d’insuline, le ratio leptine/adiponectine, le gras rétropéritonéal et le poids corporel. Ces effets ont été observés en conjonction avec une augmentation de la dépense énergétique: hausse de température à la surface de la peau (prévention) et amélioration de la fonction mitochondriale et de la synthèse d'ATP (traitement). En conclusion, l’utilisation de P. balsamifera, salicortin et L. laricina comme des traitements alternatifs et culturellement adaptés aux CEI représente une contribution importante dans la prévention et le traitement de l’obésité et du diabète.
The prevalence of obesity, insulin resistance, and the metabolic syndrome is increasing among the Cree of Eeyou Istchee (CEI - Northern Quebec). Non-traditional diet and sedentary lifestyle along with cultural disconnect of modern type 2 diabetes (T2D) therapies are involved. In order to establish culturally adapted antidiabetic treatments, our research team conducted an ethnobotanical survey, where 17 plants were identified from the CEI traditional pharmacopoeia. Based on data obtained from in vitro screening studies, two plant species out of 17 were of particular interest for their properties as antiobesity, namely Populus balsamifera L. (Salicaceae), and antidiabetic agents, namely Larix laricina K. Koch (Pinaceae). P. balsamifera and its active salicortin inhibited triglyceride accumulation during adipogenesis in 3T3-L1 adipocytes. L. laricina increased glucose uptake and AMPK activation in C2C12 myotubes, adipogenesis in the 3T3-L1 adipocyte cell line, and was observed as one of the strongest uncouplers, severely disrupting mitochondrial function (increasing fuel consumption/metabolic rate; antiobesity property). The purpose of this PhD thesis is to evaluate the antiobesity and antidiabetic potential of P. balsamifera, salicortin, and L. laricina, in an in vivo model of diet-induced obese (DIO) C57BL/6 mice, as well as to investigate their possible mechanisms of action. Mice were subjected for eight (prevention study) or sixteen weeks (treatment study) to a high fat diet (HFD), or HFD to which P. balsamifera, salicortin, or L. laricina were incorporated either at onset (prevention), or in the last 8 of the 16 weeks of administration of the HFD (treatment). The results showed that P. balsamifera (in either study) and salicortin (incorporated in HFD only in treatment study) decreased the weight of whole vii body, retroperitoneal fat pad, reduced the severity of hepatic macrovesicular steatosis and triglyceride accumulation (ERK pathway implicated). They also decreased glycemia and improved insulin sensitivity by diminishing insulin levels, and altering adipokine secretion whereby reducing the leptin/adiponectin ratio. In both studies, P. balsamifera significantly reduced food intake. This appetite-reducing effect needs to be investigated further. In the prevention study this was accompanied by an increase in energy expenditure (increase in skin temperature and tends to increase expression of uncoupling protein-1; UCP-1). The signaling pathways activated by P. balsamifera and slightly by salicortin are implicated in either controlling hepatic glucose output (Akt), skeletal muscle Glut4 expression, glucose uptake and lipid metabolism in adipose tissue (Akt), adipocyte differentiation (ERK pathway and PPARg), decreasing the hepatic inflammatory state (IKKab), and increasing muscular, hepatic, or adipose tissue fatty acid oxidation (PPARa, CPT-1). As for L. laricina, it effectively decreased glycemia levels, insulin levels and the leptin/adiponectin ratio, improved insulin sensitivity and slightly decreased abdominal fat pad and body weights. This occurred in conjunction with increased energy expenditure as demonstrated by elevated skin temperature in the prevention study, and tendency to improve mitochondrial function and ATP synthesis in the treatment protocol. In conclusion, these results represent a major contribution, identifying P. balsamifera, salicortin, and L. laricina, as promising alternative, and culturally adapted therapies for the prevention and treatment care of obesity and diabetes among the CEI.

Orientação: Maria Lídia Palma
Rumex crispus L., de nome comum Labaça-crespa, é uma planta endémica espalhada pelo mundo, sendo uma das mais infestantes em Portugal. Esta monografia, contem a descrição da planta, nomeadamente os sinónimos, variedades, o habitat e distribuição geográfica. São apresentados os extratos e a forma de obtenção. São referidos os constituintes mais importantes da planta nomeadamente, compostos fenólicos (antraquinonas, taninos, flavonoides), derivados naftalénicos, ácidos fenólicos (ácido gálico), mucilagens, o ácido oxálico, sais de ferro e fósforo. Fazem ainda parte desta monografia as informações clínicas, e principais indicações descritas na literatura (obstipação, diarreia, dermatite, urticária, psoríase, gengivite, tinha, anemia). Estudos “in vivo” e “in vitro” que suportam as ações farmacológicas estudadas (acção antimicrobiana, antioxidante, anti tumoral, anti fúngica e anti malárica).
Rumex crispus L., common name Curly dock, is a plant found throughout the world, being one of the most weeds in Portugal. This monograph contains a description of the plant, including synonyms, varieties, the habitat and geographical distribution. It presents the extracts and how they are obtained. The most important constituents of the plant, particularly phenolic compounds (anthraquinone, tannins, flavonoids), naphthalene derivatives, phenolic acids (gallic acid), mucilage, oxalic acid, salts of iron and phosphorus. They’re part of this monograph the clinical information and the main indications described in the literature (constipation, diarrhea, dermatitis, urticaria, psoriasis, gingivitis, ringworm and anemia). As well in vivo and in vitro studies support the pharmacological studied actions (antimicrobial action, antioxidant, anti tumoural, anti fungal and anti malaria).

Orientação : Maria de Fátima Frazão
As plantas foram os primeiros recursos utilizados pelos povos como parte integral da fitoterapia. A fitoterapia, etimologicamente deriva dos termos gregos therapeia (tratamento) e phyton (vegetal). Significa o estudo de plantas consideradas medicinais e da sua aplicação no tratamento de doenças. Com o passar dos anos o uso das plantas como agentes terapêuticos, contribuiu muito para o desenvolvimento de novos fármacos pelo que, tornou-se imprescindível conhecer os seus princípios activos, os mecanismos de acção subjacentes, os efeitos secundários e as possíveis interacções com os fármacos para que pudessem ser empregues de forma segura no complemento de terapias convencionais. Muitos metabolitos secundários [flavonóides] derivados de plantas são conhecidos por possuírem importantes actividades farmacológicas ao actuarem sobre o sistema biológico como antioxidantes e anti-inflamatórios. Os flavonóides estão amplamente distribuídos nas plantas como produto do seu metabolismo. Entre as diversas actividades biológicas atribuídas a estes metabolitos destacam-se as actividades antioxidantes e anti-inflamatórias, abordadas nesta monografia. Este trabalho pretende rever os extractos naturais, que contenham flavonóides com acção antioxidante e anti-inflamatória em patologias cardiovasculares e geniturinárias. Estas populações foram escolhidas pelo facto das doenças cardiovasculares serem a principal causa de morte na União Europeia, e devido à importância das doenças do trato geniturinário terem aumentado muito na medicina clínica, pois essa é uma área essencial para a compreensão de muitas doenças. Torna-se importante o enfoque de algumas dessas doenças através do presente trabalho, numa tentativa de ampliar o conhecimento sobre o assunto. Embora existam muitos estudos nesta área apenas serão consultados estudos que refiram um efeito bem documentado da acção terapêutica dos flavonóides.
Plants were the first source used by people for phytotherapy purposes. The word phytotherapy come from the greek therapeia (treatment) and phyton (vegetable). It mostly consists of the study of medicinal plants and the way its used to treatment of several diseases. Along the years the use of plants as therapeutically agents significantly contributed to the development of new pharmaceutical substances. For this reason, it became crucial to know its active components and underlying action mechanisms, secondary effects and potential interactions with other pharmaceutical substances in order to be used in a safe and effective manner for conventional therapies. Several secondary metabolites [flavonoids] derived from plants usually known to have relevant pharmaceutical impacts acting as anti oxidants and anti inflammatory on the biological system. The flavonoids are basically plant metabolites. Within many of its beneficial effects, the most relevant are anti oxidant and anti inflammatory as mentioned in this study. This paper aims to review the natural extracts containing flavonoids with antioxidant activity and anti-inflammatory genitourinary and cardiovascular action on pathologies. These populations were chosen because cardiovascular diseases are the leading cause of death in the European Union, and because of the importance of diseases of the genitourinary tract nowadays in clinical medicine, since this is a key area for understanding many diseases. It is important the focus of some of these diseases through the present work, an attempt to expand knowledge on the subject. Although there are many studies in this area only well documented studies of the therapeutic action of flavonoids will be dicussed in this monography.

South Africa is home to a wide diversity of cultural groups, all of which utilize the flora for a variety of purposes. This is true with regard to traditional medicine systems which are similar to those of the rest of Africa south of the Sahara, with diviners (sangomas) and herbalists (inyangas) as the key health providers. In addition, the Country is rich in plant diversity with some 30 000 species of flowering plants - almost one tenth of the worlds recorded higher plants. This incorporates a large diversity of plants including trees, shrubs, herbs, bulbs and corms. The adverse effects of traditional medicinal plants and natural products are not well documented in the literature. Recently, many plants used as food or in traditional medicine have been shown to be potentially mutagenic using in vitro assays. Thus, the scientific evaluation of traditional medicine and medicinal plants is very important to validate claims made on safety and efficiency of such usages. After a survey of the available ethnobotanical literature, ten trees used in South African traditional medicine were selected. These species were: Acacia niolotica subspecies kraussiana, Acacia sieberiana, Albizia adianthifolia, Combretum kraussii, Faidherbia albida, Ficus sur, Prunus africana, Salix mucronata, Terminalia sericea and Trichilia dregeana. Plant parts including leaf, root and bark were collected from each of the selected trees (exceptions were Albizia adianthifolia, Faidherbia albida, Terminalia sericea and Prunus africana) and extracted using ethyl acetate, ethanol and water individually to ensure the extraction of compounds over a wide range of polarities. The extracts (in total, 78) were screened for antibacterial, anti-inflammatory (COX-1 and COX-2) and antiacetylcholinesterase activities and investigated for their potential mutagenic effects using the Ames test. Antibacterial activity was detected using the disc-diffusion and microdilution assays. The extracts were tested against Gram-positive bacteria: Bacillus subtilis, Staphylococcus aureus, Micrococcus luteus and Gram-negative bacteria: Escherichia coli and Klebsiella pneumoniae. Of the 78 different plant extracts 111 tested (final amount of plant material was 1 mg per disc), 84% showed activity against Gram-positive bacteria. From this percentage, 20% also showed activity against Gram-negative bacteria. The best inhibition was observed with ethyl acetate and ethanol root extracts of Terminalia sericea against both Gram-positive and Gram-negative bacteria. In the micro-dilution assay, 55% of the plant extracts showed minimum inhibitory concentration (MIC) values ~ 1.56 mg/ml against Gram-positive and/or Gram-negative bacteria. The ethyl acetate bark extract of Acacia sieberiana and the root and bark ethyl acetate extracts of Acacia nilotica inhibited bacterial growth of both Gram-positive and Gram-negative bacteria at concentrations ~ 0.8 mg/ml. The aqueous leaf extracts of Acacia sieberiana had a low MIC value (0.3 mg/ml) against Gram-negative Kleibsiella pneumoniae and the ethyl acetate extracts of the root inhibited growth of Escherichia coli with an MIC value of 0.1 mg/ml. However, these two extracts showed no activity in the disc-diffusion assay. The MIC values of the neomycin (control) were 0.8 I-Ig/ml and 3.1 I-Ig/ml against Kleibsiella pneumoniae and Escherichia coli respectively. In the anti-inflammatory test, 70% of the plant extracts from different plant parts (leaf, root, bark) of the tree investigated showed strong inhibition in both the CQX-1 and CQX-2 bioassays. The CQX-2 inhibitory effects of aqueous extracts were generally lower when compared to the organic solvent extracts. However, water extracts of Acacia nilotica was an exception (~ 90%). In the acetylcholinesterase inhibitory test, 21% of the plant extracts were active at concentrations ~ 1 mg/ml using the micro-plate assay. The lowest IC50 value was 0.04 mg/ml obtained with an ethanol bark extract of Combretum kraussii. The IC50 value of the galanthamine (positive control) was 2 I-IM. None of the investigated plants showed any potential mutagenic effects with Salmonella typhymurium strain TA 98 using the Ames test. Using bioassay-guided fractionation, anolignan B was isolated from the ethyl acetate root extract of Terminalia sericea. Antibacterial activity of anolignan B was determined using the microdilution assay. The compound possessed activity against both Gram-positive and Gram-negative bacteria. The lowest MIC value (3.8 IJg/ml) was observed with Staphylococcus aureus. MIC value of the neomycin was 1.5 IJg/ml. Anti-inflammatory activity of anolignan B was detected using the CQX-1 and CQX-2 bioasays. The compound showed strong inhibitory activity against CQX-1 and weaker activity against CQX-2. The ICso values were 1.5 mM and 7.5 mM with CQX-1 and CQX-2 respectively. The ICso values of indomethacin were 0.003 mM and 0.186 mM against CQX-1 and CQX-2 respectively. There were no potential mutagenic effects showen by anolignan B against Salmonella typhimurium strain TA 98 in the Ames test. Isolation of anolignan B from Terminalia species and the antibacterial and anti-inflammatory activities observed in this work have not been reported previously and could therefore be recorded as novel biological activities for this compound. These results also support the idea that the use of ethnobotanical data can provide a valuable short cut by indicating plants with specific uses which might likely be sources of biologically active chemicals.
Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2005.

Nous avons utilisé une approche ethnobotanique pour identifier des espèces de plantes utilisées par les Cris afin de traiter les symptômes du diabète de type 2. Larix laricina du Roi (L. laricina) a récemment été identifiée comme une des meilleures plantes qui a stimulé le transport de glucose dans les cellules C2C12 et fortement potentialisé la différenciation des 3T3-L1 en indiquant une sensibilité potentiellement accrue à l’insuline. Ensuite, ces études de criblage ont été effectuées sur des extraits éthanolique (EE) en utilisant une série de bioessais in vitro. Cependant, les préparations traditionnelles des plantes sont souvent faites avec l’eau chaude. Le but de cette thèse de doctorat était d’isoler les principes actifs de L. laricina par un fractionnement guidé par l’adipogenèse; d’évaluer et de comparer l’activité et les mécanismes antidiabétiques des EE et des extraits aqueux (HWE) de ces 17 plantes. Pour le fractionnement de L. laricina, on a isolé plusieurs composés connus et identifié un nouveau composé actif cycloartane triterpene, qui a amélioré fortement l’adipogenèse et a été responsable en partie de l’activité adipogénique (potentiellement similaire à l’effet sensibilisateur à l’insuline des glitazone) de l’extrait éthanolique issu de l’écorce de L. laricina. Pour le métabolisme lipidique, nos résultats ont confirmé que 10 parmi les 17 EE ont augmenté la différenciation des adipocytes alors que 2 extraits seulement l’ont inhibée. Les HWE ont montré une faible activité adipogénique ou antiadipogénique. Les EE de R. groenlandicum et K. angustifolia ont le PPAR γ (peroxisome proliferator-activated receptor γ), le SREBP-1 (sterol regulatory element binding protein-1) et le C/EBP (CCAAT-enhancer binding proteins) α, alors que ceux de P. balsamifera et A. incana les ont inhibés. L’effet inhibiteur de P. balsamifera a également été prouvé d’avoir impliqué l’activation de la protéine kinase activée par l’AMP (AMPK). Les EE et HWE de R. groenlandicum ont stimulé les mêmes facteurs de transcription alors que les extraits aqueux d’autres plantes sélectionnées ont perdu ces effets en comparaison avec leurs extraits éthanoliques respectifs. L’analyse phytochimique a également identifié le groupe des espèces actives et inactives, notamment lorsque les espèces ont été séparées par famille de plante. Finalement concernant l’homéostasie de glucose, nos résultats ont confirmé que plusieurs EE ont stimulé le transport de glucose musculaire et inhibé l’activité de la glucose-6-phosphatase (G6Pase) hépatique. Certains des HWE ont partiellement ou complètement perdu ces activités antidiabétiques par rapport aux EE, tandis qu’une seule plante (R.groenlandicum) a juste conservé un potentiel similaire entre les EE et HWE dans les deux essais. Dans les cellules musculaires, les EE de R.groenlandicum, A. incana et S. purpurea ont stimulé le transport de glucose en activant la voie de signalisation de l’AMPK et en augmentant le niveau d’expression des GLUT4. En comparaison avec les EE, les HWE de R.groenlandicum ont montré des activités similaires; les HWE de A. incana ont complètement perdu leur effet sur tous les paramètres étudiés; les HWE de S. purpurea ont activé la voie de l’insuline au lieu de celle de l’AMPK pour augmenter le transport de glucose. Dans les cellules H4IIE, les EE et HWE des 5 plantes ont activé la voie de l’AMPK, et en plus les EE et HWE de 2 plantes ont activé la voie de l’insuline. La quercétine-3-O-galactoside et la quercétine 3-O-α-L-arabinopyranoside ont été identifiées comme des composés ayant un fort potentiel antidiabétique et donc responsables de l'activité biologique des plantes HWE actifs avec le transport du glucose. En conclusion, on a isolé plusieurs composés connus et identifié un nouveau triterpène actif à partir du fractionnement de L. laricina. Nous avons fourni également une preuve directe pour l'évaluation et la comparaison d'une action analogue à l'insuline ou insulino-sensibilisateur des EE et HWE de plantes médicinales Cris au niveau de muscle, de foie et de tissus adipeux. Une partie de leur action peut être liée à la stimulation des voies de signalisation intracellulaire insulino-dépendante et non-insulino-dépendante, ainsi que l’activation de PPARγ. Nos résultats indiquent que les espèces de plantes, les tissus ou les cellules cibles, ainsi que les méthodes d'extraction sont tous des déterminants significatifs de l'activité biologique de plantes médicinales Cris sur le métabolisme glucidique et lipidique.
We have used a collaborative ethnobotanical approach to identify plant species used by the Cree of Eeyou Istchee (CEI) to treat symptoms of type 2 diabetes. Several screening studies were performed on 17 species identified in a survey of the Cree Nation. Firstly, Larix laricina du Roi (L. laricina) was recently identified as one of the top plants, which stimulated glucose uptake in C2C12 muscle cells and strongly potentiated the differentiation of 3T3-L1 pre-adipocytes suggesting enhanced insulin sensitivity. Secondly, these screening studies were performed on ethanol extracts (EE) using an in vitro bioassay platform, however, traditional preparations are often based on hot water. So the purpose of this PhD thesis was to isolate the active principles from L. laricina through adipogenesis-guided fractionation, and to evaluate and compare the antidiabetic activity and mechanisms of EE and hot water extracts (HWE) of these 17 Cree plants. For the fractionation of L. laricina, we isolated several known compounds and identified a new active cycloartane triterpene, which strongly enhanced adipogenesis in 3T3-L1 cells and was responsible partly for the adipogenic (potentially glitazone-like insulin sensitizing) activity of the ethanol extract of the bark of L. laricina. In the adipocyte lipid metabolism course, the results confirmed that 10 of the 17 EE stimulated adipocyte differentiation and adipogenesis, whereas 2 had inhibitory effects. Corresponding HWE exhibited partial or complete loss of such adipogenic or anti-adipogenic activity. R. groenlandicum and K. angustifolia EEs activated Peroxisome proliferator-activated receptor γ (PPAR γ), sterol regulatory element binding protein-1 (SREBP-1) and CCAAT-enhancer binding protein (C/EBP) α, whereas P. balsamifera and A. incana decreased these transcription factors. P. balsamifera’s inhibitory effect was also found to involve AMP-activated protein kinase (AMPK) activation. R. groenlandicum HWE and EE stimulated similar transcription factors, but HWE of other selected plants lost such effects compared to their respective EE. Phytochemical analysis also uncovered clustering of active versus inactive species, notably when species were segregated by plant family. The results showed that several EE stimulated muscle glucose uptake and inhibited hepatic glucose-6-phosphatase (G6Pase) activity. Some of the HWE partially or completely lost these antidiabetic activities in comparison to EE; while one plant (R.groenlandicum) retained similar potential between EE and HWE in both assays. In C2C12 muscle cells, EE of R.groenlandicum, A. incana and S. purpurea stimulated glucose uptake by activating AMPK pathway and increasing GLUT4 expression level. In comparison to EE, HWE of R.groenlandicum exhibited similar activities; HWE of A. incana completely lost its effect on all parameters; interestingly, HWE of S. purpurea activated insulin pathway instead of AMPK pathway to increase glucose uptake. In the H4IIE cells, all selected 5 plants HWE and EE activated AMPK pathway, and in addition, 2 plants EE and HWE also activated insulin pathways. Quercetin-3-O-galactoside and quercetin 3-O-α-L-arabinopyranoside were identified as potential candidates to be responsible for the biological activity of the active HWE plants in the glucose transport assay. In conclusion, we isolated several known compounds and identified a new active triterpene from fractionation of L. laricina. We also provide direct evidence evaluating and comparing of an insulin-like or insulin-sensitizing action of EE and HWE of Cree medicinal plants at the level of muscle, liver and adipose tissue. Part of their actions may be related to stimulation of insulin-dependent and insulin-independent intracellular signaling pathways, as well as to PPARγ activation. The results indicate that plant species, target tissues or cells, as well as extraction methods, are all significant determinants of the biological activity of Cree medicinal plants on glucose and lipid metabolism.

Orientação: Patrícia Rijo
As plantas constituem um recurso terapêutico ainda hoje muito utilizado. O interesse mundial no uso das plantas medicinais tem sido crescente, com os seus efeitos benéficos a serem redescobertos para o desenvolvimento de novos medicamentos. Pela primeira vez, com esta dissertação, foi elaborada uma revisão bibliográfica sistemática com o objectivo de clarificar o estado actual dos estudos sobre Rosmarinus officinalis L., elucidando quais os seus compostos e as actividades biológicas mais relevantes. Para tal, foi feita uma pesquisa na base de dados ‘PUBMED’, com os termos “rosemary” e “Rosmarinus officinalis”, donde foram recolhidos 232 artigos publicados entre 1990 e 30 de Novembro de 2014 sobre as actividades farmacológicas e os compostos isolados do alecrim. Ao longo dos anos verificou-se um crescente interesse nas propriedades desta planta. O ácido carnósico, o óleo essencial, o carnosol e o ácido rosmarínico foram os compostos mais estudados, e as actividades anti-tumoral, anti-infecciosa e antioxidante as mais reportadas. Pela consistência de estudos, o ácido carnósico e o óleo essencial de R. officinalis, demonstram ser alternativas prometedoras para o desenvolvimento de novos fármacos antitumorais e anti-infecciosos, respectivamente. Com esta revisão, conclui-se que o alecrim constitui uma excelente fonte de compostos com inúmeras propriedades medicinais.
Plants are a therapeutic resource still widely used nowadays. The worldwide interest in the use of medicinal plants has been growing, with its beneficial effects being rediscovered for the development of new drugs. For the first time, with this work, it´s presented a systematic literature review in order to clarify the current state of research on Rosmarinus officinalis L., elucidating which compounds and biological activities are the most relevant. To this end, a search was made in the database PUBMED, with the terms "rosemary" and "Rosmarinus officinalis", which included 232 articles published between 1990 and November 30, 2014 about rosemary’s pharmacological activities and their isolated compounds. Over the years there has been an increasing interest in the properties of this plant. The carnosic acid, essential oil, carnosol, and rosmarinic acid were the most studied compounds and antitumoral, anti-infective and antioxidante the most activities reported. By the consistency of studies, carnosic acid and R. officinalis essential oil, shown to be promising alternatives for the development of new anti-tumoral and anti-infective drugs, respectively. With this review, we conclude that rosemary is an excellent source of compounds with numerous medicinal properties.

Orientação: Patrícia Rijo
O uso de produtos à base de plantas para o tratamento, cura e prevenção de doenças, constitui uma das mais antigas práticas medicinais da humanidade. De facto, uma fracção significativa das populações dos países em desenvolvimento permanece dependente dos conhecimentos ancestrais sobre plantas para os seus cuidados de saúde. Ainda assim, continua a existir uma lacuna entre os progressos observados na farmácia clínica e os no campo da fitoterapia e da medicina tradicional, continuando muitos produtos naturais com actividade biológica por identificar. As espécies de Plectranthus (família Lamiaceae) têm uma generalizada aplicação etnobotânica, sendo frequentemente citadas as suas propriedades medicinais e aplicação, sobretudo ao nível da medicina popular. A composição rica em compostos antioxidantes e os diversos efeitos apresentados (anti-inflamatório, antimicrobiano e antifúngico), sugerem o Plectranthus como um promissor género para a descoberta de compostos medicinais. Assim, o isolamento dos metabolitos secundários das espécies Plectranthus é importante para validar cientificamente os usos populares dessas plantas e também para encontrar novas fontes de produtos com potencial económico, ou compostos que possam ser transformados em princípios activos. Além disso, a avaliação da citotoxicidade dos extractos de plantas e os seus princípios activos são necessários para uma utilização terapêutica eficaz e segura. Nesta dissertação enumeram-se os 28 compostos isolados até hoje nos extractos da espécie Plectranthus ecklonii Benth. e respectivas bioactividades. Os resultados da análise HPLC apresentados fazem parte de um projecto, actualmente a decorrer no CBIOS, sobre identificação, quantificação e avaliação de compostos presentes em diferentes espécies de Plectranthus, nomeadamente diterpenos e ácidos hidrocinâmicos.
The use of herbal products for the treatment, prevention and cure of diseases is one of the oldest human medicinal practices. In fact, a main fraction of population in developing countries remains dependent on ancestral plant knowledge for health care. However, there‘s still a gap between progress observed in clinical pharmacy and in the field of herbal and traditional medicine, remaining many natural products with biological activity to be identified. Plectranthus species (Lamiaceae family) have a widespread ethnobotanical use and are often cited by its medicinal properties and application, particularly in folk medicine. They contain many antioxidant compounds and exhibit several effects (anti-inflammatory, antimicrobial and antifungal) which suggest that Plectranthus may be a promising genus for the discovery of medicinal compounds. Thus, the isolation of secondary metabolite compounds from the Plectranthus spp. is important to validate scientifically the popular uses of these plants and also to find new sources of potentially economically important products or compounds which can be transformed into active pharmaceutical ingredients. Besides, the cytotoxicity evaluation of the plant extracts and their active ingredients are required for their effective and safe therapeutic use. This dissertation enumerates the 28 compounds isolated to date from Plectranthus ecklonii Benth., extracts and their biological activities. The HPLC analysis presented is part of a project currently ongoing at CBIOS, of identification, quantification and evaluation of the bioactive components, in particular diterpenes and hydrocinnamic acids, in different species of Plectranthus.