Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2017-09-05T21:41:06Z No. of bitstreams: 1 PolyannaSilvaMoreira_DISSERT.pdf: 1108101 bytes, checksum: 99f0f4dcef670d145f1ddf9eb582a862 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2017-09-18T21:33:56Z (GMT) No. of bitstreams: 1 PolyannaSilvaMoreira_DISSERT.pdf: 1108101 bytes, checksum: 99f0f4dcef670d145f1ddf9eb582a862 (MD5) / Made available in DSpace on 2017-09-18T21:33:56Z (GMT). No. of bitstreams: 1 PolyannaSilvaMoreira_DISSERT.pdf: 1108101 bytes, checksum: 99f0f4dcef670d145f1ddf9eb582a862 (MD5) Previous issue date: 2017-02-23 / A dengue ? uma arbovirose que afeta o homem, gerando uma problem?tica na sa?de p?blica do mundo, especialmente em pa?ses tropicais os quais apresentam condi??es que favorecem a dissemina??o do mosquito Aedes aegypti. Atualmente, dentre as v?rias estrat?gias para controle da doen?a, ainda n?o se tem uma vacina eficaz ou um antiviral capaz de combater essa infec??o. Assim, o objetivo do presente estudo foi avaliar a atividade antiviral de extratos obtidos da folha e frutos da planta Morinda citrifolia L. em cultura de c?lulas Vero infectadas com v?rus dengue-2 (DENV-2). Inicialmente foram obtidos os extratos brutos (hidroetan?lico) e as respectivas fra??es: hexano, clorof?rmio e acetato de etila, analisados por cromatografia. O teste de citotoxicidade do extrato bruto, res?duo aquoso e fra??es foram realizados em cultura de c?lulas Vero pelo m?todo MTT, nas concentra??es de 1000; 500; 250; 125; 62,5; 31,2 ?g/mL. O ensaio antiviral foi conduzido atrav?s das seguintes estrat?gias: c?lulas infectadas com DENV-2 (controle positivo); c?lulas mantidas com meio de cultura (controle negativo); c?lulas infectadas com DENV-2 e tratadas com o extrato ou fra??es. Ap?s cinco dias de infec??o a viabilidade celular foi avaliada pelo m?todo de MTT e o sobrenadante da cultura foi utilizado para quantifica??o viral por unidade formadora de placa (PFU). Os resultados demonstraram que a an?lise cromatogr?fica dos extratos e fra??es revelou bandas distintas e sugestivas de saponinas, terpenos e flavonoides. Tais extratos e fra??es n?o foram t?xicos para as culturas de c?lulas, com exce??o do tratamento das c?lulas com a fra??o clorof?rmio obtido da folha e as fra??es hexano e acetato de etila do fruto verde, levando a uma viabilidade pr?xima de 65%. No ensaio antiviral o controle positivo apresentou viabilidade celular em torno de 60% ap?s cinco dias de infec??o. No tratamento com os compostos obtidos da folha observou-se que ao adicionar a fra??o de acetato de etila ?s c?lulas infectadas, estas mantiveram uma viabilidade celular pr?ximo a 100% na concentra??o de 1000?g/mL e a 85% nas concentra??es de 500 e 250?g/mL. O tratamento com a fra??o hexano apresentou uma viabilidade superior ao controle positivo em todas as concentra??es. No entanto, na fra??o clorof?rmio, a viabilidade manteve-se elevada apenas nas concentra??es de 500 e 250?g/mL. O extrato bruto e a fra??o residual aquosa n?o demonstraram atividade antiviral. As c?lulas tratadas com o extrato e as diferentes fra??es obtidas dos frutos maduro e verde, apresentaram de um modo geral uma viabilidade celular pr?xima de 100% nas concentra??es de 500 e 1000 ?g/mL no fruto maduro e apenas 1000 ?g/mL no fruto verde, com exce??o das c?lulas que foram tratadas com a fra??o clorof?rmio, na qual n?o foi poss?vel observar nenhuma diferen?a significativa quando comparado ao controle positivo. Na quantifica??o viral observou-se que as c?lulas tratadas com as fra??es hexano e clorof?rmio obtidos da folha e tamb?m os extratos brutos obtidos dos frutos maduro e verde tiveram a??o antiviral, resultando na diminui??o total da carga viral. Finalmente, a partir desse estudo podemos identificar uma poss?vel atividade antiviral dos compostos obtidos de Morinda citrifolia contra o v?rus dengue. / Dengue is an arbovirosis which affects mankind, causing problems in the public health worldwide, especially in tropical countries which present conditions that favor the spread of the mosquito Aedes aegypti. Currently, among the various strategies to control the disease, there is no effective vaccine or antiviral capable of combating this infection. Thus, the aim of the present study was to evaluate the antiviral activity of leaf and fruit extracts of the plant Morinda citrifolia L. in Vero cells culture infected with dengue-2 virus (DENV-2). Initially, the crude extracts (hydroethanolic) and their fractions were obtained: hexane, chloroform and ethyl acetate, followed by chromatographic analysis. The cytotoxicity test of the crude extract, the aqueous residue and its fractions were performed in culture of Vero cells by the MTT method, at concentrations of 1000; 500; 250; 125; 62.5; 31.2 ?g/mL. The antiviral assay results were conducted through the following strategies: cells infected with DENV-2 (positive control); cells maintained with culture medium (negative control); cells infected with DENV-2 and treated with the extract or fractions. After five days of infection, cell viability was evaluated by the MTT method and culture supernatant was used for viral quantification by plaque forming unit (PFU) assay. The results showed that the chromatographic analysis of extracts and fractions present distinct bands, which could be suggestive of saponins, terpenes and flavonoids. Such extracts and fractions were not toxic to cell cultures, except for the cells treated with the chloroform fraction obtained from leaf, hexane and ethyl acetate fractions of the green fruit, leading to a near 65% viability. In the antiviral assay the positive control had 60% of cell viability after five days of infection. Among the leaf extract treatments, it was observed that infected cells treated with ethyl acetate fraction, maintained their cell viability around 100% in the concentration of 1000?g/mL and up to 85% in the concentrations of 500 and 250?g/mL. The hexane fraction treatment showed higher viability in comparison to the positive control at all concentrations. However, in the chloroform fraction, viability remained high only at concentrations of 500 and 250 ?g/mL. Crude extract and residual aqueous fraction did not show any antiviral activity. Cells treated with the extract and different fractions obtained from the mature and green fruits, presented an overall cell viability close to 100% in 500 and 1000 ?g/mL in the mature fruit and only 1000 ?g/mL in the green fruit. However, for the cells treated with the chloroform fraction, it was not possible to observe any significant difference when compared to the positive control. In the viral quantification it was observed that cells treated with hexane and chloroform fractions obtained from leaf, as well as crude extracts obtained from mature and green fruits had antiviral effect, resulting in a total viral load decrease. Finally, identified from this study, a possible antiviral activity of the compounds obtained from Morinda citrifolia against dengue virus.

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Syftet med den här studien är att undersöka hur rörlig bild kan öka konsumentens förtroende för en produkt vid e-handel, och för att se om det eventuella ökandet av förtroende för produkter kan få fler kunder att vilja handla online. Målet med studien är att se hur påverkad användaren blir av rörlig bild när den handlar kläder på en webbplats. Den utvalda metoden för studien var en observation med sju kvinnor i form av ett användartest där respondenterna fick visualisera ett köp av en specifik produkt på två olika webbplatser, med en efterföljande intervju med tio öppna frågor. Resultatet av studien visade att majoriteten av de tillfrågade föredrog att handla på en webbplats som använde rörlig bild vid presentation av produkten, eftersom det gav en mer realistisk bild av klänningen. Slutsatsen resulterade i att rörlig bild visualiserade produkten mer verklighetstroget till skillnad från en e-handelsplats som endast använde produktbilder. Den rörliga bilden var det närmaste konsumenten kunde komma till verkligenheten och det ökade i sin tur förtroendet för produkten och har resulterat i att fler konsumenter ville handla produkten. Den rörliga bilden fungerade därmed som ett substitut för den fysiska kontakten konsumenten annars skulle haft med produkten i realiteten. Slutsatsen kan därför dras om att rörlig bild ökar förtroendet för en specifik produkt vid e-handel. / The purpose of the study is to research how moving images can increase the confidence for the customer in a specific product in e-commerce. Furthermore, the study aimed to see if moving images could increase the number of people wanting to shop online. The reason behind the study is to see how customers who are interested in buying clothes respond to moving images. The chosen method for the study was to observe seven women participating in a usability test to envision purchasing a chosen product on two separate websites. Following the usability test, each woman was interviewed with ten open questions.The results of the study showed that the majority of the respondents preferred to shop on awebsite that used moving images to present a product, as it presented the product in a morerealistic image. Because the respondents could not see the dress in reality, a catwalk video was the most effective way to present the dress to the customer. The catwalk video thus increased the confidence of the customer in the product, in which subsequently lead to an increased number of consumers wanting to purchase the product. The moving images served as a substitute for the physical contact the consumer would otherwise have with the product in reality. It is therefore appropriate to conclude that moving images can increase the confidence in a customer for a certain product in e-commerce.

In this study, Naegleria fowleri, a pathogenic amoeba and the causative agent of Primary Amebic Meningoencephalitis (PAM), was utilized to determine the presence or absence of classically conserved Golgi molecules featured in the expression of a Golgi apparatus. Previous studies concluded no Golgi expression via light microscopy and transmission electron microscopy, but a recent report on Naegleria gruberi indicated the presence of dispersed Golgi tubules. Non-pathogenic species of the Naegleria genus such as Naegleria gruberi 30540 and Naegleria lovaniensis 30569 were utilized in Western immunoblot analysis compared to reduced whole-cell lysate proteins of two strains of N. fowleri and Vero CCL-81, Chlorocebus sp. kidney epithelial cells, which were utilized as a positive control for Golgi expression. N. fowleri and N. lovaniensis whole-cell lysates had indications of a 110 kDa reduced protein, associated with the predicted molecular weights of the beta-COPI subunit of the COPI cis-Golgi vesicular transport complex with further Western immunoblot indication of a weak band around 25 kDa corresponding to rabbit polyclonal antibodies specific for ARF1. Serial Dilutions of Wheat Germ Agglutinin Alexa Fluor 488TM were performed on Vero cells, Naegleria fowleri 30894, and N. gruberi 30540 with 1:100 dilution of recommended stock dilution of WGA 488 determined for utilization in sequential immunofluorescence. Sequential immunofluorescence with Wheat Germ Agglutinin Alexa Fluor 488TM and then blocked with 3% BSA:PBS [wt/vol] dilution with subsequent incubation in rabbit anti-beta-COPI primary 1:250, and 1:1000 of Alexa Fluor 594 goat anti-rabbit secondary antibody exposure showed strong indications of organized cis- and trans-punctate Golgi body markers in close association in individual and dividing cells of Naegleria fowleri and conserved Golgi expression in the positive control Vero cells, but further experiments are necessary to verify this finding with N. fowleri.

MSc (Microbiology) / Department of Microbiology / Background: The medicinal value of plants lies in some chemical substances that produce a definite physiological action in the human body. The secondary metabolites help the plants to survive hash conditions and could be used by humans as supplements of their health, as foods additives or for medicinal purposes. This bioactive compounds are not always beneficial to human beings, and some of this plants bioactive compounds can be toxic or genotoxic to human cells. This study used several methods to evaluate of phytochemical constituents and mutagenic properties of Coccinia rehmanni and Jatropha zeyheri plant extracts. Methodology: Methanol was used for extraction of the bioactive compounds from the two selected plants, filtered with Whatman filter paper and evaporated with rotary evaporator. The extracts were fractionated using open column chromatography. Chemical and TLC methods were used to determine phytochemicals of the study plants extracts and fractions. The plants extracts and fractions were tested against Vero cell lines in order to evaluate cytotoxicity and genotoxicity of the plants. NucRed and LTR Hoechst 33342 dyes were used for cytotoxicity and genotoxicity respectively. For the evaluation of cytotoxicity and genotoxicity Quantification of live and dead cells for the screening assay was performed using the ImageXpress Micro XLS Widefield Microscope and acquired images analyses using the MetaXpress software and Multi-Wavelength Cell Scoring Application Module. Antimutagenicity of plants extracts was observed using PARP universal colorimetric assay kit. Acquired data was transferred to an EXCEL spreadsheet and data was analyzed. Results and discussion: C. rehmanni (12.03%) yielded more extract than J. Zeyheri (8.20%). the two plants had different compound composition and were in different stages of maturity. The study revealed the domination of Terpenoids, Cardiac glycosides, Phenolic and tannis. With an exception of two fraction fractions all the fractions was found to be toxic to an extent were genotoxicity of such fraction could not be concluded. The reason for such extreme toxicity could be due to the influence of the retained alcohol during rotary evaporation. xvi | P a g e Conclusion: this study provides and add to existing knowledge on the phytochemicals mutagenicity and anti-mutagenicity of C. rehmanni and J. Zeyheri medicinal plants. The study serves as scientific proof that extensive use of this plant in traditional medicine for treatment of various ailments may lead to some irreversible damages. / NRF

Guatemalan medicinal plants were collected and screened for the presence of essential oils using steam distillation. Oil was found in 63 species from 24 families and was tested in tube dilution assays for activity against Escherichia coli, Staphylococcus aureus, Streptococcus mutans, Lactobacillus acidophilus and Candida albicans. Several essential oils were highly active with 20 instances of oils inhibiting the microbes at an MIC of 0.31 µl/ml. Oils were also tested against cancerous and established cell lines using a 15% (v/v) agar-media which was developed to improve essential oil solubility. Assays were performed against three cancer lines: Stomach (AGS: CRL-1739), Skin (A375: CRL-1619), Tongue (CAL27: CRL-2095) and an established Monkey Kidney cell line (Vero C 1008: CRL-1586). Assessment of viability was performed using the Neutral Red assay with results indicating that many of the oils significantly inhibited cancer cell lines in vitro with 24 individual instances producing an IC50 of 0.20 µl/ml or less. Therapeutic indices indicated that many of the highly inhibitory oils were more cytotoxic to cancerous cell lines than to the established cell line.

No description available.

Ce travail de thèse s'attache à identifier des plantes et/ou molécules à activités cytotoxique, antioxydante, anti-inflammatoire et antivirale ciblant le virus du chikungunya (CHIKV) dans le but de trouver des alternatives thérapeutiques vis-à-vis du stress oxydatif et de l'inflammation, mécanismes impliqués dans les maladies chroniques non transmissibles (diabète, obésité…), et de la maladie du chikungunya, maladie vectorielle réémergente. La première partie de ces travaux présente les résultats obtenus lors d'un criblage d'activités biologiques réalisé sur une sélection de dix-huit plantes endémiques et indigènes de La Réunion. Les activités ciblées ont été les activités cytotoxiques sur une lignée cellulaire humaine (cellules THP-1), les activités antioxydantes évaluées par un test in cellulo d'hémolyse et par quatre tests chimiques (TEAC/DPPH/FRAP/ORAC) ainsi qu'une évaluation de la teneur en composés phénoliques (test FOLIN) et les activités anti-inflammatoires testées sur des macrophages murins (cellules RAW-BlueTM). Les résultats obtenus ont permis de mettre, plus particulièrement, en évidence les activités de différents extraits : cytotoxique pour Carissa spinarum, antioxydantes pour Agarista buxifolia et Dryopteris wallichiana et anti-inflammatoire pour Stillingia lineata et Indigofera ammoxylum. La deuxième partie du travail est consacrée à l'étude phytochimique d'une espèce indigène de La Réunion, Stillingia lineata, choisie en raison des résultats obtenus lors de ce criblage biologique préliminaire et de ceux du programme Phytochik. Un fractionnement bioguidé par un test antiviral, réalisé sur des cellules Vero (cellules rénales de singe vert Cercopithecus aethiops) contaminées par le CHIKV, a conduit à l'isolement de trois macrocycles diterpéniques rares de type tonantzitlolone dont l'un présente une structure non caractérisée jusque-là, et d'un pimarane de structure nouvelle. La 4'-acétoxytonantzitlolone a été identifiée comme molécule candidate contre le CHIKV (CE50 = 7 μM). Des relations structure-activité ont pu être définies ; la présence d'un groupement oxygéné sur la chaîne latérale des tonantzitlolones semble jouer un rôle important sur la réponse antivirale de ces squelettes diterpéniques. / The aims of this PhD work were to identify plants and/or molecules with cytotoxic, antioxidant, anti-inflammatory or antiviral (chikungunya virus , CHIKV) activities in order to find therapeutic alternatives towards oxidative stress and inflammation, mechanisms involved in chronic noncommunicable diseases (diabetes, obesity ...), and chikungunya disease, reemerging vector-borne disease. The first part of this work presents the results obtained from a biological screening carried out on a selection of eighteen endemic and indigenous plants of La Réunion. The targeted activities were cytotoxicity on a human cell line (THP-1), antioxidant activities evaluated using an in cellulo hemolysis assay and four chemical tests (TEAC / DPPH / FRAP / ORAC) together with an evaluation of the content of phenolic compounds (FOLIN test) and anti-inflammatory activity tested in murine macrophages (RAW cells-BlueTM). The results allowed to highlight activities of different extracts in particular : cytotoxic for Carissa spinarum, antioxidant for Dryopteris wallichiana and Agarista buxifolia and anti-inflammatory for Stillingia lineata and Indigofera ammoxylum.The second part of this work is devoted to the phytochemical study of Stillingia lineata, an indigenous species of La Réunion chosen because of the results obtained in this preliminary biological screening and those carried out in Phytochik programme. Bioassay-guided fractionation performed on Vero cells (green monkey kidney cells Cercopithecus aethiops) infected with CHIKV led to the isolation of three rare macrocycle-type diterpenes called tonantzitlolone and a new pimarane. The 4'-acetoxytonantzitlolone was identified as a candidate molecule against CHIKV (EC50 = 7 μM). Structure-activity relationships have been defined, the presence of an oxygenated group on the side chain of tonantzitlolones seems to play an important role in the antiviral response of the diterpene skeleton.

<p>Management of predation on sea turtle nesting beaches is vital to conservation efforts for the vulnerable loggerhead turtles (<i>Caretta caretta</i>) and olive ridley turtles (<i>Lepidochelys olivacea</i>). Sea turtles increasingly face threats from invasive and human-tolerant mammalian predators as human disturbances on nesting beaches rises. The intensity of mammalian predation has increased in Las Baulas National Park in Costa Rica which is an important nesting site for several species of threatened and endangered sea turtles. I analyzed loggerhead and olive ridley nest predation on four beaches in the United States and Costa Rica that were chosen for variations in degree of human disturbance and management strategies. My objectives were to 1) determine if egg predation rates differ at the four sites, 2) determine the most destructive predators at each location, and 3) suggest management options to alleviate mammalian threats to turtle clutches on Playa Grande and Playa Cabuyal in Costa Rica. My results show that the beaches without a nest protection or predator control program had very high rates of predation. Invasive mammalian predators and mammalian predators associated with human disturbance were the most destructive at the four sites. I recommend that regulations regarding dogs and the take of eggs from the beach are enforced at Playa Cabuyal and that physical nest protection is rapidly implemented at Playa Grande. I also recommend that the National Park consider managing raccoon predation by removing problem individuals, but caution that they do so in a way that maintains the animals’ role in the ecosystem.</p>

In Günter Grass's Danzig Trilogy and novella, Im Krebsgang, an antagonistic female type makes a repeated appearance. She appears in the guise of Susi Kater and Luzie Rennwand in Die Blechtrommel, and as Tulla Pokriefke in the other works, Katz und Maus, Hundejahre, and Im Krebsgang. This antagonistic female type is not like other women in these works. A review of Le Deuxième Sexe by feminist Simone de Beauvoir reveals several crucial components contributing to woman's position in society. Most essentially, a woman's natural attributes and (dis)abilities and the conventions of society have enforced her historical submission to man. This thesis analyzes how the antagonistic female type, or villain, compares and contrasts with other female figures in these works by Grass, according to a paradigm derived from Beauvoir's description of woman. From this analysis, a better understanding of the female villain's nature emerges. Indeed, such a comparison demonstrates that certain female figures in the works of Grass transcend their historically oppressed or subdued status by refusing to submit to those natural handicaps and societal restrictions identified by Beauvoir, and thus become a threat to man's status or security as an antagonistic female type, or villain. However, the villain figure is not always inherently evil, but possesses the capacity to change. The villain and victim can reconcile their differences and may even form a friendly relationship. This evolving villain-victim duality becomes most clear in Grass's work, Im Krebsgang, and suggests the possibility of assuaging contemporary conflicts as educators sympathize with the experiences of both extremist groups and victimized parties and help them come to terms with their differences.