L. japonicus, o popular \"rubim\", pertence à família Lamiaceae. O presente trabalho tem como objetivo contribuir e complementar trabalho realizado anteriormente sobre a mesma espécie, em relação à caracterização farmacobotânica, aspectos químicos e farmacológicos, incluindo atividade antiúlcera, antioxidante e antimicrobiana. A droga, constituída de folhas e caule, foi caracterizada macro e microscopicamente. O extrato hidroetanólico 70% liofilizado (EHEL), preparado com as partes aéreas, obtido através de percolação, apresentou na triagem fitoquímica presença de flavonóides, taninos, saponinas e 0,06% de óleos essenciais. Os taninos presentes na droga vegetal e no extrato foram quantificados e os teores foram respectivamente de 0,06% e de 0,35%. O teor de flavonóides na droga vegetal foi de 0,76% e no EHEL, de 2,3%. O extrato foi fracionado por solventes de polaridades diferentes como clorofórmio, acetato de etila, etanol 50% e etanol. Foi determinado o perfil cromatográfico para o extrato e frações. O extrato apresentou atividade antiúlcera extremamente significativa, no modelo de indução por etanol acidificado, administrado na dose de 400 mg/kg, por via oral, reduzindo índice de Lesão, Área Total de Lesão e Área Relativa de Lesão, sendo mais evidente no próprio extrato e nas frações clorofórmica e acetato e etila, comparativamente ao controle. Não houve inibição de crescimento de Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Campylobacter coli em uma concentração de até 2.000 µg/mL, verificado através da determinação da concentração mínima inibitória pelo método de difusão em placa. O extrato não apresentou resultado significativo em relação a atividade antioxidante testada através do método que reduz o radical 2,2\'-difenil-1-picrilhidrazil (DPPH), e após o equilíbrio da reação, permite calcular a quantidade de antioxidante gasta para reduzir 50% do DPPH. / L. japonicus, popularly known as \"rubim\", belongs to the Lamiaceae family. The objective of the present study is to contribute and complement previous investigations on the same species, in relation to pharmacobotanical characterization, chemical and pharmacological aspects, including antiulcer, antioxidant e antibiotic activities. The drug, formed of leaves and stalk, was macro e microscopically qualified. Flavonoids, tannins, saponins and 0.06% of essential oils are present in the lyophilized 70% hydroalcoholic extract (HE), obtained through percolation. Tannins are present at 0,06% in the vegetable drug and at 0,35% in the extract. The content of flavonoids in the drug was at 0,76% and in the extract (HE), at 2,3%. The extract was fractioned by solvents of different polarities as chloroform, ethyl acetate, absolute ethanol and 50% ethanol. The chromatographic profile of the extract and fractions was determined. The extract, orally administered at the dose of 400 mg/kg, presented extremely significant antiulcer activity in the acidified ethanol induction model, by reduction of the Index of Lesion, Total Area of Lesion and Relative Area of Lesion. The values of the extract and of chloroformic and ethyl acetate fractions were most evident, in relation to control. There was no inhibition of growth of Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Campylobacter coli at a concentration up to 2.000 µg/mL, verified through the the minimum inhibitory concentration by the plate diffusion method. The extract didn\'t present significant result in relation to antioxidant activity tested through the method that reduces the radical 2,2\'-Diphenyl-1-picrylhydrazyl (DPPH), and after the balance of the reaction, it allows to calculate the amount of antioxidant necessary to reduce 50% of DPPH.

Espécies do gênero Copaifera (Leguminosae) são nativas de regiões tropicais da América Latina e África. No Brasil, seu óleo-resina é amplamente utilizado em medicina popular como antiinflamatório, antisséptico e cicatrizante. No presente trabalho, avaliou-se, em uma primeira fase, a atividade antiinflamatória em modelo experimental agudo das seguintes amostras de óleo-resina: comercial, gentilmente cedida pela empresa Pronatus, Copaifera reticulata, C. multijuga e C. paupera. As amostras foram administradas pela via oral, sendo selecionada, das amostras identificadas botanicamente, a C. reticulata por apresentarem maior atividade. A amostra comercial também foi ensaiada no modelo experimental acima. Os óleo-resinas foram analisados por cromatografia gasosa acoplada à espectrometria de massas tendo revelado a presença de sesquiterpenos e diterpenos. Foram identificados vinte e dois compostos na amostra comercial e vinte seis na amostra de C. reticulata. Deste total, seis substâncias foram comuns a ambas as amostras. A seguir, os óleos-resinas foram destilados e separadas suas respectivas frações sesqui e diterpênicas. Tanto os óleos-resinas quanto suas frações foram avaliados farmacologicamente. As atividades farmacológicas testadas foram: atividade antiinflamatória aguda, subcrônica, crônica e ensaios de toxicidade aguda, subcrônica e citotoxicidade. A atividade antiinflamatória foi testada em cinco modelos: edema de pata induzido pela carragenina, nistatina e miconazol; indução de formação de tecido granulomatoso e dermatite induzida pelo óleo de cróton. Nos modelos utilizados as doses testadas foram de 2,47 ml/kg (DE50 no modelo da carragenina) para a amostra comercial e suas frações sesqui e diterpênicas e 2,02 ml/kg (Dose equipotente no modelo da carragenina) para a amostra de C. reticulata e suas frações sesqui e diterpênicas). Em todos os ensaios realizados com as amostras, evidenciaram-se intensa atividade antiflogística, com exceção da C. reticulata que apresentou elevada toxicidade no modelo de indução do tecido granulomatoso. Nos ensaios de toxicidade aguda, a amostra C. reticulata (DL50 = 4,48 ml/kg) apresentou toxicidade maior que a amostra comercial (DL50 > 12,35 ml/kg). Suas frações sesquiterpênicas foram bem menos tóxicas que seus correspondentes óleos-resinas com DL50 maiores que 9,14 e 20 ml/kg, respectivamente para C. reticulata e amostra comercial: Por outro lado, a fração resinosa (diterpênica) apresentou alta toxicidade com DL50 superiores a 2,85 e 1,29 ml/kg, respectivamente para a amostra comercial e C. reticulata. Nos ensaios de toxicidade subcrônica confirmou-se a toxicidade menor para a fração sesquiterpênica da amostra de C. reticulata. O óleo-resina de C. reticulata apresentou toxicidade renal e alterações hepáticas indicando que pode haver um comprometimento hepático. / Species of the genus Copaifera (Leguminosae) are native of tropical regions in Latin America and Africa. In Brazil, its oil-resin is widely utilized in popular medicine as anti-inflammatory, antiseptic and cicatrizing medicine. In the present work, we initially evaluated the acute anti-inflammatory activity of the following samples: commercial, Copaifera reticulata, C. multijuga and C. paupera. All the samples were administered by oral route. This initial pharmacological evaluation revealed that the C. reticulata was clearly more effective. In the second part of this work, the C. reticulata and the commercial samples, kindly provided by Pronatus-Manaus, were investigated. The oil-resins of the selected samples were analyzed by gas chromatography connected with mass spectrometry, revealing the presence of sesquiterpens and diterpens. Twenty two compounds in the commercial sample and twenty six in the C. reticulata were identified. Then after, the oil-resins were distilled and separated in their respective sesqui and diterpens fractions. Both oil-resins and their fractions were pharmacologically evaluated. The pharmacological evaluations were: acute, sub-chronic and chronic anti-inflammatory activities and acute, sub-chronic toxicities and cito-toxicities assays. The anti-inflammatory activity was studied in five experimental models: carrageenan, nistatin and miconazole paw oedema; induction of granulomatous tissue formation and croton oil dermatitis. The administered doses by oral route were 2.47 ml/kg (EQ50 for carrageenan model) to commercial sample and its sesqui and diterpens fractions, and 2.02 ml/kg to C. reticulata sample and its sesqui and diterpens fractions. All the assays carried out with the selected samples showed intense anti-flogistic activity, except for C. reticulata that exhibited high toxicity in the granulomatous tissue model. In the acute toxicity studies, C. reticulata (LD50 = 4.48 ml/kg) revealed greater toxicity in comparison with the commercial sample (LD50 > 12.35 ml/kg). Its sesquiterpens fractions were less toxic that its correspondent oil-resins with LD50 higher than 9.14 and 20 m1/kg, for C. reticulata and commercial sample, respectively. On the other hand, resin fraction (diterpenic) presented high toxicity with LD50 higher than 2.85 and 1.29 ml/kg for commercial and C. reticulata samples, respectively. The sub-chronic assays, confirmed the less toxicity for the C. reticulta sesquiterpenic fraction. The C. reticulata oil-resin presented renal toxicity and hepatics alterations.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / A Aroeira do SertÃo (Myracrodruon urundeuva AllemÃo) à uma planta medicinal utilizada popularmente no tratamento de problemas ginecolÃgicos, tais como Cervicite, Vaginites e Ectopias, cuja forma farmacÃutica de creme vaginal à de 15%; utilizada a mais de uma dÃcada pelo Programa FarmÃcias Vivas. Objetivou-se estudar o perfil de utilizaÃÃo e a eficÃcia do creme de Aroeira em ginecologia; notificar e quantificar possÃveis reaÃÃes adversas a fitoterÃpicos. Trata-se de estudo observacional, prospectivo, de tipo aberto e nÃo comparativo de utilizaÃÃo de medicamentos. Desenvolvido em um Centro de SaÃde, pertencente à regional IV de Fortaleza-CE. O estudo foi composto por 33 mulheres que receberam indicaÃÃo e prescriÃÃo de uso do creme vaginal de Aroeira, sendo que apenas 18 completaram todas as fases do estudo. Os dados foram coletados entre janeiro e junho de 2008, mediante acompanhamento de mulheres que concluÃram o estudo. Utilizou-se como tÃcnica a entrevista individual prà e pÃs-tratamento; a reuniÃo de informaÃÃes clÃnicas fornecidas pela ginecologista responsÃvel, alÃm de um formulÃrio para registro dos dados clÃnicos. Os resultados demonstraram que nenhuma reaÃÃo adversa foi relatada e o creme mostrou-se efetivo no tratamento das patologias diagnosticadas (Cervicites e Ectopias). Conclui-se, assim, que os achados corroboram todos os efeitos farmacolÃgicos jà detectados em relaÃÃo à aroeira e que esta nÃo acarreta nenhum tipo de reaÃÃo adversa, confirmando, dessa forma, o que preconiza as diretrizes da nova PolÃtica Nacional de Plantas Medicinais, voltadas para a saÃde da mulher. / The Brazilian Peppertree (Myracrodruon urundeuva AllemÃo) is a medicinal plant popularly known in the treatment of gynecological problems, such as cervicitis, vaginitis and ectopies, which pharmaceutical form in cream is 15%, used for more than ten years by the Live Pharmacy Program. One aimed to study the use and efficiency of Brazilian Peppertree in gynecology; record and quantify possible adverse and phytotherapic reactions. It is an observatory, prospective study, of the open type and non comparative of medicine use. Developed in a Health Unit, from regional IV in Fortaleza-CE. The study was composed by 33 women that received indication and prescription of Brazilian Peppertreeâs vaginal cream, but only 18 completed all the phases of the study. The data were collected between January and June 2008, by accompaniment of women who concluded the study. One used as technique the individual interview pre and post treatment; the use of clinical information given by the responsible gynecologist, besides a form to record the clinical data. The results demonstrated that no adverse reaction was reported and that the cream was effective in the treatment of the diseases (Cervicitis and Ectopies). One concludes, thus, that the findings confirm all the pharmacological effects already detected concerning the Brazilian Peppertree and that it does not cause any adverse reaction, confirming, this way, what preaches the new National Policy of Medicinal Plants, regarding womenâs health.

No presente trabalho, realizou-se o estudo químico da fração alcaloídica total de Duguetia lanceolata St. Hil. (Annonaceae), espécie proveniente do cerrado paulista, tendo-se obtido o perfil cromatográfico e a separação decomponentes da fração alcaloídica, que resultou na obtenção do alcaloide aporfínico majoritário N-acetilanonaína, isolado da espécie, pela primeira vez. No estudo botânico de folhas, foram descritas as características da morfologia externa e interna, tendo sido ilustradas por fotografias e fotomicrografias dos itens mais importantes para a diagnose da droga caracteres xeromórficos, como: consistência coriácea, cutícula espessa, escamas e tricomas tectores estrelares nas epidermes, presença de bainhas e grandes grupos de células esclerenquimáticas freqüentes, na nervura mediana e no pecíolo. Entre outros elementos de destaque encontram-se: a distribuição de drusas na epiderme e a presença de células oleíferas dispersas no mesofilo. A avaliação da atividade antiprotozoária mostrou, de forma geral, que frente ao Plasmodium falciparum, os alcalóides totais são mais ativos sobre a cepa resistente à cloroquina (K-1) (CE50:2,0µg/mL) o que também foi observado para os mesmos componentes acetilados (CE50:3,0µg/mL). Tanto o extrato etanólico, quanto os alcaloides totais (l00µg/mL) da espécie, demonstraram elevados níveis de atividade frente às formas tripomastigotas de Tripanosoma cruzi, tendo apresentado percentual de morte de cerca de 100% e 66%, respectivamente. Este último, foi comparável ao benznidazol, fármaco usado como referência. Tais resultados são promissores tendo em vista que, os extratos não apresentaram citotoxicidade sobre as células de camundongos (RAW 264.7), até a maior concentração testada (120µg/mL). No ensaio de citotoxicidade sobre Artemia salina, os extratos etanólico e alcaloídico foram ambos inativos (DL50>1000µg/mL), em contraste com os alcalóides totais acetilados que demonstraram nível de toxicidade comparável ao do sulfato de atropina (DL50:867,60µg/mL) utilizado como referência, no ensaio. / ln this work, some chemical, botanical and pharmacological aspects of the brazilian Duguetia lanceolate St. Hil (Annonaceae) were considered. The plant specimen was collected from the cerrado biome, at São Paulo, Brazil. The isoquinoline alkaloid fraction was extracted and then acetylated. After performed the chromatographic profile, the column fractionation led to the first isolation of the major aporphine N- acetyl-anonaine from this specie. Morphological and anatomical characters of the leaves were described and illustrated by photographies and photomicrographies. Some xeromorphic characters, typical from cerrado species, were recognized: coriaceous leaves, thick cuticle, scales, stellate non-glandular trichomes, sclerenchymatic vascular sheats and scattered sclerenchymatic cell groups in the median nervure and petiole. Other important features were observed: isolated druses of calcium oxalate in each epidermal cells and oil cells scattered in the mesophyll. Evaluation of the antiprotozoal activity showed that the total alkaloid fraction (EC50:2,0µg/mL) and the same acetylated fraction (EC50:3,0µg/mL), were more active against the chloroquine resistant strain (K-1) of Plasmodium falciparum. The total acetylated alkaloids and the ethanolic extract (l00µg/mL) were highly active (0% of survival) against trypomastigotes of Tripanosoma cruzi. ln the same test, non acetylated alkaloids (l00µg/mL) killed about 66% of the parasite forms, having an activity comparable to Benznidazole, the standard drug. Those results were promising, considering the low cytoxicity showed by the plant extracts (ethanol and total alkaloid) over mice macrophages RAW 264.7, at the maximum tested concentration (120 µg/mL). The ethanol and total alkaloid extracts showed, also, low levels of citotoxicity (LD50>l000µg/mL), when assayed against the Artemia salina while the acetylated alkaloids had a LD50 value (843,2 µg/ml) comparable to the atropine sulphate.

Não consta resumo na publicação. / The objective of the present dissertation is to contribute for the enlargment and engrossment of the available data on the drug known as maracujá as well as on its extracts. Comprised topics include comparative morphodiagnostic studies and illustrations for P.alata Dryand., P. quadrangularis L., P. edulis Sims. and P. incarnata L.. The differentiation and characterization of the above mentioned species in terms of chemical composition was accomplished by thin layer chromatographic techniques. Chromatographical profiles were established for flavonoid and alkaloid fractions on the four species in relation to reference standards orientin, isoorientin, vitexin, and isovitexin (for flavonoids), and harman and harmin (for alkaloids). P. alata Dryand. earned a more detailed study. The quantifications of flavonoid and alkaloid fractions were performed densitometricaly by TLC and expressed as orientin and harman, respectively. Pharmacological assays carried out with extracts included acute toxicity and CNS depressor activity.

The relationship between the endocannabinoid system in the renal medulla and the long-term regulation of blood pressure is not well understood. To investigate the possible role of the endocannabinoid system in renomedullary interstitial cells, mouse medullary interstitial cells (MMICs) were obtained, cultured and characterized for their responses to treatment with a selective inhibitor of fatty acid amide hydrolase (FAAH), PF-3845. Treatment of MMICs with PF-3845 increased cytoplasmic lipid granules detected by Sudan Black B staining and multilamellar bodies identified by transmission electron microscopy. HPLC analyses of lipid extracts of MMIC culture medium revealed a 205nm-absorbing peak that showed responsiveness to PF-3845 treatment. The biologic activities of the PF-3845-induced product (PIP) isolated by HPLC were investigated in anesthetized, normotensive surgically-instrumented mice. Intramedullary and intravenous infusion of PIP at low dose rates (0.5-1 AU/10 min) stimulated diuresis and natriuresis, whereas at higher doses, these parameters returned toward baseline but mean arterial pressure (MAP) was lowered. Whereas intravenous bolus doses of PIP stimulated diuresis, GFR and medullary blood flow (MBF) and reduced or had no effect on MAP, an intraperitoneal bolus injection of PIP reduced MAP, increased MBF, and had no effect on urinary parameters. Genetic or pharmacological ablation of the cannabinoid type 1 receptors in mice completely abolished the diuretic and vasodepressor properties of intramedullary infused PIP, suggesting that the PF-3845-induced product requires the presence of CB1 receptors in order to elicit its renal effects. In a radioactive competition binding assay, using Chinese hamster ovary cells expressing CB1 receptors, PIP successfully displaced the CB1 selective inverse agonist [3H] SR141716A, revealing that the lipid extract was able to compete for binding to CB1 receptors. Finally, we investigated the tubular location of diuretic activity that the PF-3845-induced lipid fraction exhibits. In a renal function in vivo experiment, we pre-treated anesthetized mice with an intramedullary infusion of one of four well-known diuretics. This procedure was followed by an intramedullary infusion of PIP (1AU). Only inhibition of the proximal tubule sodium reabsorption diminished the diuretic activity of the PF-3845-induced product, suggesting that the lipid fraction requires a physiologically intact proximal tubular reabsorption mechanism for it to produce diuresis. These data support a model whereby PF-3845 treatment of MMICs results in increased secretion of a neutral lipid which acts directly to promote diuresis and natriuresis and indirectly through metabolites to produce vasodepression. Efforts to identify the structure of the PF-3845-induced lipid and its relationship to the previously proposed renomedullary antihypertensive lipids are ongoing.

Background This thesis investigated the efficacy and safety of Bacopa monnieri in improving memory in healthy Australians over the age of 55-years. A review of the literature showed that memory impairment and dementia are increasingly prevalent in the current demographic climate of an ageing population. As well as the pathological cognitive loss of neurodegenerative disease, many older persons are experiencing memory loss as part of the physiological process of ageing. Bacopa monnieri is a herbal medicine used since antiquity in the traditional Ayurvedic medical system of India for its cognitive enhancing effects. A number of pre-clinical and clinical studies support this traditional usage. Laboratory studies have demonstrated antioxidant and cholinergic actions in the brain as well as improved memory and cognitive performance in animal models. Human trials of Bacopa have also demonstrated improved memory performance. Some of these trials are limited by methodological flaws such as lack of blinding, small sample sizes, or use of outcome measurements which are not well validated. However, a small number of well designed human trials provide evidence for efficacy in cognitive and memory performance improvement. The current study was employed to extend on previous findings by assessing the efficacy and safety of Bacopa in the aged population specifically, as it is in this population that memory impairment becomes apparent. Aims 1. To assess the efficacy of Bacopa monnieri in improving memory in healthy Australians over the age of 55-years. 2. To assess whether the use of Bacopa is associated with side-effects Design A 12-week, randomised, double-blind, placebo-controlled, parallel group clinical trial. Participants Participants were self selected from the general population. They were aged 55-years or over at the commencement of the trial. Participants were without dementia, depression or other serious health conditions and did not use psychotropic medications. Intervention Participants were randomised to one of two treatment conditions, either a tableted extract of Bacopa monnieri called Bacomind™ (300mg/day, standardised to contain at least 40% bacosides), or an identical placebo. Participants attended three clinical evaluations: the first an initial screening session, the second a baseline evaluation of neuropsychological function and subjective memory performance at the commencement of the trial and the third, an end-of-trial outcome evaluation at 12-weeks, during which neuropsychological function and subjective memory performance were again assessed along with side-effects and study compliance. Primary Outcome Measures Rey Auditory Verbal Learning Test (AVLT), Rey-Osterrieth Complex Figure Test (CFT), Memory Complaint Questionnaire (MAC-Q), and Trail Making Test (TMT) Results From 136 people who elected to participate, 103 people met study entry criteria and 98 of these commenced the trial. Of these, 81 participants completed the trial and provided evaluable data for the end point analysis. Bacopa monnieri versus placebo significantly improved verbal learning as well as delayed recall as measured by the AVLT (p<.05). Though improvements were noted in the CFT, MAC-Q and TMT, there were no significant differences between placebo and active groups found for these tests. The Bacopa group reported a higher incidence of gastro-intestinal (GIT) side-effects than the placebo group, these predominantly being increased stool frequency, abdominal cramps and nausea. No other significant adverse effects were found. Conclusions A clinical trial was carried out to assess the effects of 12-weeks administration of Bacopa monnieri (300mg/day) on memory performance in people over the age of 55-years. Primary outcome measures were well validated neuropsychological tests that objectively measured verbal and visual memory and a memory complaint questionnaire that measured subjective memory complaints. The results demonstrated that Bacopa significantly improved memory acquisition and retention in older Australians. This concurs with findings from previous human and animal studies, as well as supports traditional Ayurvedic claims and uses. The beneficial effects on memory observed may be due to previously demonstrated antioxidant and cholinergic effects of the herb on the central nervous system. The use of Bacopa was associated with GIT side-effects, particularly increased bowel movements, nausea and abdominal cramping, findings infrequently reported previously. Possible explanations for these side-effects include GIT irritation by the saponin constituents of the herb, or cholinergic stimulation of autonomic and motor responses in the GIT, or a combination of both of these factors. The side-effects observed in the current study provide supportive evidence that Bacopa may increase cholinergic activity in humans. A worthwhile future extension of the current study would be to assess whether the finding of Bacopa’s efficacy for improving memory performance is replicable in populations with either mild cognitive impairment or early dementia.

<p>Marine invertebrates, in particular sponges, represent a source of a wide range of secondary metabolites, many of which have been attributed various defensive capabilities against environmental stress factors. In this thesis sponge-derived low-molecular peptide-like compounds and associated analogs are investigated for bioactivity and pharmacological targets. </p><p>The compound bromobenzisoxazolone barettin (cyclo[(6-bromo-8-(6-bromo-benzioxazol -3(1H)-one)-8-hydroxy)tryptophan)]arginine) was isolated from the sponge <i>Geodia barretti</i> and its ability to inhibit larval settlement of the barnacle <i>Balanus improvisus</i> was determined. With an EC₅₀ value of 15 nM, this compound’s antifouling effect was higher than those of the previously reported brominated dipeptides from <i>Geodia barretti</i>, i.e., barettin and 8,9-dihydrobarettin; moreover, this antifouling effect was demonstrated to be reversible. However, the compound lacked affinity for 5-HT_1-7 receptors, whereas barettin possessed specific affinity to 5-HT_2A, 5-HT_2C and 5-HT₄, while 8,9-dihydrobarettin interacted with 5-HT₄. In an attempt to evaluate structure-activity relationships synthesized analogs with barettin and dipodazine scaffolds were investigated for antifouling activity. The analog benso[g]dipodazine, with an EC₅₀ value of 34 nM, displayed the highest settlement inhibition.</p><p>The studies of the structure-activity relationships of sponge-derived compounds were extended to cover analogs of agelasines and agelasimines originally isolated from sponges of the genus <i>Agelas</i>. Synthesized (+)-agelasine D and two structurally close analogs were investigated for cytotoxic and antibacterial activity. The profound cytotoxicity and broad spectrum antibacterial activity found prompted a further investigation of structure-activity relationships in 42 agelasine and agelasimine analogs and several characteristics that increased bioactivity were identified.</p><p>In conclusion this work has produced new results regarding the potent bioactivity of compounds derived from the sponges <i>Geodia barretti</i> and <i>Agelas</i> spp. and increased SAR knowledge of the fouling inhibition, cytotoxicity and antimicrobial activity of these compounds.</p>

Soy contains many bioactive molecules known to elicit anti-cancer effects. One such peptide, Lunasin, has been shown to selectively act on newly transformed cells while having no cytotoxic effect on non-tumorigenic or established cancer cell lines. In this study we attempt to understand the developmental effects of Lunasin overexpression in vivo and create reagents that will help us understand Lunasin’s anti tumorigenic effects in an intact organism. cDNA encoding lunasin and EGFP-lunasin were cloned into pUAST and microinjected into Drosophila embryos. Tissue-specific overexpression of EGFP-Lun in the resulting transgenic lines was accomplished by crossing transgenics to various GAL4 driver lines. Progeny were assessed for phenotypic alterations and no phenotypic abnormalities were observed in tissues expressing EGFP-Lunasin, supporting current studies that show Lunasin does not affect normal cells. Previous studies have localized Lunasin to the nuclear compartment. To test if this was the case for EGFP-Lun, subcellular localization of EGFP-Lun was determined via fluorescence microscopy. Salivary glands from EGFP-Lun expressing individuals were dissected, fixed, and mounted in Vectashield® with the nuclear stain, DAPI. Our results demonstrate that EGFP-Lun, like native Lunasin, is localized to the nucleus. Eight transgenic lines were mapped to specific chromosomes and EGFP-Lun transgenic line GEJ1-L2 was balanced in preparation for use in tumor suppression studies. In summary, we have created and characterized transgenic flies capable of overexpressing Lunasin under the control of the GAL4/UAS system. Localization of EGFP-Lunasin to the nucleus and data on the phenotypic consequence of its overexpression in flies is presented. Finally, reagents created as part of this thesis will aid experiments aimed at understanding the effects of Lunasin on benign and invasive tumors.

D. Tech. Pharmaceutical Sciences. / Aims of the research project was to develop and optimise rapid quality control methods for H. gordonii raw material and products. The second aim was to determine whether the perceived active component of H. gordonii (P57) is transported across porcine intestinal and buccal mucosa.