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Prospecção de bioativos farmacológicos em algas marinhas Rhodophyta e Heterokontophyta e avaliação de citotoxicidade / Prospection of pharmaceutical bioactive compounds in marine algae Rhodophyta and Heterokontophyta and cytotoxicity evaluation.Daniel Xavier Andreguetti 15 October 2015 (has links)
Muitas drogas terapêuticas produzidas pela indústria farmacêutica são estruturas químicas isoladas de organismos encontrados na natureza ou moléculas baseadas nelas. Podem ser incluídas nesse grupo drogas isoladas de organismos marinhos, como corais, esponjas e algas marinhas, conhecidos como produtores de grandes quantidades de metabólitos secundários. Com base neste fato o presente estudo teve como objetivo realizar a prospecção de moléculas bioativas com propósito farmacológico, em extratos de algas marinhas vermelhas (Rhodophyta) e pardas (Heterokontophyta) coletadas no litoral brasileiro. A prospecção foi realizada por meio de avaliação de seus potenciais antioxidante, antibacteriano, antifúngico, anticancerígeno, e antiparasitário contra organismos causadores de leishmaniose e esquistossomose. Para as avaliações foram empregadas os extratos supercríticos de 5 espécies diferentes, sendo 2 pardas: Dictyota dichotoma e D. menstrualis e três vermelhas: Chondria littoralis, Spyridia hypnoides e Plocamium brasiliense. Os extratos foram avaliados quanto aos seus potenciais bioativos e os resultados mais promissores foram selecionados para as etapas seguintes do fracionamento. Em uma avaliação geral os extratos apresentaram bons resultados e representam uma potencial fonte de bioativos. Os extratos das espécies de D. dichotoma e D. menstrualis foram então submetidos a um procedimento de fracionamento bioguiado pela atividade esquistossomicida. Incorporou-se ainda um terceiro extrato de D. mertensii aos estudos e todas as etapas do fracionamento foram monitoradas por LC-MS. Comparando-se as massas detectadas em todas as frações que apresentaram atividade, para os 3 extratos, foi verificado que a substância de m/z 271,24 estava presente em todas elas, portanto os procedimentos de isolamento foram direcionados a esta molécula para a qual foi possível isolar 7 mg. Diferentemente do que era esperado a molécula quando avaliada isoladamente não apresentou atividade esquistossomicida, levando a hipótese de que a atividade seja decorrente de uma molécula diferente para cada espécie ou ainda que a mesma seja decorrente de uma interação com outras substâncias por um mecanismo de ação aditivo ou sinérgico. O trabalho avaliado de forma geral apresentou resultados promissores e representa um grande embasamento para servir como base para posteriores trabalho de fracionamento. / Several therapeutic drugs manufactured by the pharmaceutical industry are chemical structures isolated from organisms that are found in nature or molecules based on that. May be included at this group drugs isolated from marine organisms, like corals, sponges and seaweeds, known as great secondary metabolites producers. Based on this facts the objective of the present study is to perform a prospection study to achieve bioactive molecules with pharmaceutical purposes, on extracts made from red (Rhodophyta) and brown (Heterokontophyta) seaweed collected in the Brazilian shore. The prospection studies was performed by means of evaluation of the antioxidant, antibacterial, antifungal, anticancer and antiparasitic (against Leishmania and Schistosoma) potential. In the evaluation were tested the supercritical extracts of 5 different species, including 2 brown species: Dictyota dichotoma and D. menstrualis and 3 red species: Chondria littoralis, Spyridia hypnoides and Plocamium brasiliense. The extracts were evaluated by their potential bioactive compounds and the most promising results were selected for the following fractionation steps. Overall the extracts have shown good results and may be represent a potential source of bioactive molecules. The extracts of both D. dichotoma and D. menstrualis were submitted to a bioguided fractionation process by their antischistosomal activities. It was still included a third extract from D. mertensii to the studies and every step was monitored by LC-MS techniques. Comparing the detected mass for each active fraction, it was observed the presence of a substance with m/z 271,24 in all of the extracts, so the isolating procedures were directed to obtain that specific molecule, which was obtained in a biomass of 7 mg. Differently than expected the molecule when evaluated isolated do not show the antischistosomal activity, leading to the hypothesis that the activity was related to different molecules for each species or even the observed effect is resulted by an interaction mechanism with another substances by an additive or synergist mechanism. The overall evaluation of the whole work show some promising results and it represent a great support for future fractionation works.
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Prospecção de bioativos farmacológicos em algas marinhas Rhodophyta e Heterokontophyta e avaliação de citotoxicidade / Prospection of pharmaceutical bioactive compounds in marine algae Rhodophyta and Heterokontophyta and cytotoxicity evaluation.Andreguetti, Daniel Xavier 15 October 2015 (has links)
Muitas drogas terapêuticas produzidas pela indústria farmacêutica são estruturas químicas isoladas de organismos encontrados na natureza ou moléculas baseadas nelas. Podem ser incluídas nesse grupo drogas isoladas de organismos marinhos, como corais, esponjas e algas marinhas, conhecidos como produtores de grandes quantidades de metabólitos secundários. Com base neste fato o presente estudo teve como objetivo realizar a prospecção de moléculas bioativas com propósito farmacológico, em extratos de algas marinhas vermelhas (Rhodophyta) e pardas (Heterokontophyta) coletadas no litoral brasileiro. A prospecção foi realizada por meio de avaliação de seus potenciais antioxidante, antibacteriano, antifúngico, anticancerígeno, e antiparasitário contra organismos causadores de leishmaniose e esquistossomose. Para as avaliações foram empregadas os extratos supercríticos de 5 espécies diferentes, sendo 2 pardas: Dictyota dichotoma e D. menstrualis e três vermelhas: Chondria littoralis, Spyridia hypnoides e Plocamium brasiliense. Os extratos foram avaliados quanto aos seus potenciais bioativos e os resultados mais promissores foram selecionados para as etapas seguintes do fracionamento. Em uma avaliação geral os extratos apresentaram bons resultados e representam uma potencial fonte de bioativos. Os extratos das espécies de D. dichotoma e D. menstrualis foram então submetidos a um procedimento de fracionamento bioguiado pela atividade esquistossomicida. Incorporou-se ainda um terceiro extrato de D. mertensii aos estudos e todas as etapas do fracionamento foram monitoradas por LC-MS. Comparando-se as massas detectadas em todas as frações que apresentaram atividade, para os 3 extratos, foi verificado que a substância de m/z 271,24 estava presente em todas elas, portanto os procedimentos de isolamento foram direcionados a esta molécula para a qual foi possível isolar 7 mg. Diferentemente do que era esperado a molécula quando avaliada isoladamente não apresentou atividade esquistossomicida, levando a hipótese de que a atividade seja decorrente de uma molécula diferente para cada espécie ou ainda que a mesma seja decorrente de uma interação com outras substâncias por um mecanismo de ação aditivo ou sinérgico. O trabalho avaliado de forma geral apresentou resultados promissores e representa um grande embasamento para servir como base para posteriores trabalho de fracionamento. / Several therapeutic drugs manufactured by the pharmaceutical industry are chemical structures isolated from organisms that are found in nature or molecules based on that. May be included at this group drugs isolated from marine organisms, like corals, sponges and seaweeds, known as great secondary metabolites producers. Based on this facts the objective of the present study is to perform a prospection study to achieve bioactive molecules with pharmaceutical purposes, on extracts made from red (Rhodophyta) and brown (Heterokontophyta) seaweed collected in the Brazilian shore. The prospection studies was performed by means of evaluation of the antioxidant, antibacterial, antifungal, anticancer and antiparasitic (against Leishmania and Schistosoma) potential. In the evaluation were tested the supercritical extracts of 5 different species, including 2 brown species: Dictyota dichotoma and D. menstrualis and 3 red species: Chondria littoralis, Spyridia hypnoides and Plocamium brasiliense. The extracts were evaluated by their potential bioactive compounds and the most promising results were selected for the following fractionation steps. Overall the extracts have shown good results and may be represent a potential source of bioactive molecules. The extracts of both D. dichotoma and D. menstrualis were submitted to a bioguided fractionation process by their antischistosomal activities. It was still included a third extract from D. mertensii to the studies and every step was monitored by LC-MS techniques. Comparing the detected mass for each active fraction, it was observed the presence of a substance with m/z 271,24 in all of the extracts, so the isolating procedures were directed to obtain that specific molecule, which was obtained in a biomass of 7 mg. Differently than expected the molecule when evaluated isolated do not show the antischistosomal activity, leading to the hypothesis that the activity was related to different molecules for each species or even the observed effect is resulted by an interaction mechanism with another substances by an additive or synergist mechanism. The overall evaluation of the whole work show some promising results and it represent a great support for future fractionation works.
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Estudio de los sistemas de polisacáridos del alga parda Dictyota dichotoma y su actividad antiviralRabanal Atalaya, Melissa 27 April 2015 (has links)
Los objetivos iniciales de este trabajo de tesis consistieron en la extracción de los polisacáridos de la feofita Dictyota dichotoma, el fraccionamiento y la elucidación estructural de los polisacáridos purificados, y la evaluación de sus actividades antivirales contra los virus CVB3 y HHV1.
El primer polisacárido extraído fue el fucoidano. El alga seca y molida se extrajo con etanol a temperatura ambiente y a 70 °C; seguidamente se extrajo con HCl diluido a temperatura ambiente y a 70 °C, por nueve veces consecutivamente. Sobre el residuo se extrajo el alginato con Na<sub>2</sub>CO<sub>3</sub> al 3% a 50 °C. Los polisacáridos provenientes de la extracción con HCl se fraccionaron a través de la precipitación con Cetrimide y posterior redisolución con concentraciones crecientes de 0,5, 1, 1,5, 2, 4 y en algunos casos de 5 M de NaCl. El estudio estructural se basó en la metilación de los polisacáridos fraccionados y en la espectroscopía de resonancia magnética nuclear bidimensional, particularmente HSQC. Además, se realizaron análisis de las fracciones purificadas sobre los virus de CVB3 y HHV1. El fucoidano extraído a temperatura ambiente (EAR) mostró características de un polisacárido heterodisperso con respecto a la composición de monosacáridos, siendo la fucosa el principal monosacárido (36%), seguido de galactosa (23%), y proporciones más chicas de xilosa y manosa (≤ 18%). Además, dicho extracto mostró altas proporciones de hidratos de carbono (48%), ácidos urónicos (21%), medianas proporciones de grupos sulfato (15%) y un bajo peso molecular (6,5 KDa). Los extractos obtenidos a mayor temperatura también mostraron heterodispersión con respecto a la composición de monosacáridos, observando que conforme incrementaba la secuencia extractiva la cantidad de fucosa y galactosa disminuían, mientras que la cantidad de xilosa y manosa se incrementaban. Los análisis generales nos mostraron en todos los extractos altas proporciones de hidratos de carbono excepto en EAH9. Además, la cantidad de proteínas solubles y peso molecular y grupos sulfato disminuyeron, mientras que los ácidos urónicos incrementaban conforme se incrementaba la secuencia extractiva. El alginato extraído exhibió una preponderancia del ácido manurónico por sobre el ácido gulurónico. Además, los análisis mostraron que parecen triadas de dihos ácidos urónicos. El fraccionamiento con Cetrimide mostró que EAR y EAH1 tienen características de un típico galactofucano sulfatado, con cantidades muy pequeñas de otras estructuras. Conforme el fraccionamiento continúaba, a bajas redisoluciones, la cantidad de fucosa disminuía y la xilosa, manosa y ácidos urónicos se incrementaban. A medida que el fraccionamiento continúó esta tendencia predominó, la fucosa dejó de ser el monosacárido principal en la mayoría de las subfracciones en los siguientes extractos, mientras, que los manosa, xilosa y ácidos urónicos se incrementaron.
El análisis estructural de las fracciones purificadas mostraron un core basado en residuos de →2)-Man-(1→ con sustituciones en O-3 o en O-6 y →6)-Gal-(1→, sulfatada en O-3. Además, se observó que los residuos de fucosa se encontraron enlazados en proporciones variables en C-2, C-3 y C-4, y en buena medida terminales, así como altas proporciones de ramificaciones de β-D-xilosa. La acción antiviral contra el virus HHV1 en las diferentes fracciones tanto a temperatura ambiente como a 70 °C, mostró que la fracciones -0,5, con características de uronofucoidanos, presentaron la más alta actividad antiviral, seguida de las fracciones EAR-2 y EAR-4. Por otro lado, a pesar que la actividad de las fracciones obtenidas a temperatura poseían menor actividad antiviral, las fracciones EAH2-0,5, EAH4-0,5 y EAH9-0 presentaron actividades moderadas. Con respecto al virus CVB3, los extractos de la feofita D. dichotoma mostraron actividades moderadas frente este virus, siendo las fracciones EAR-1,5, EAR-4 y EAH1-4, las que presentaron la mejor actividad antiviral. Cabe señalar que la fracción EAR-4, caracterizada por presentar un fucogalactano sulfatado, mostró ser la de mayor actividad frente a ambos virus, mientras que los altos valores obtenidos de CC<sub>50</sub> en todas las fracciones indican una baja citotoxicidad para los fucoidanos extraídos de Dictyota dichotoma.
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Atividades anticoagulante e antioxidante de extratos brutos ricos em polissacar?deos sulfatados das macroalgas marinhas marrons Canistrocarpus cervicornis, Dictyota mertensii e Dictyopteris delicatula e de Heterofucanas de Canistrocarpus cervicornisC?mara, Rafael Barros Gomes da 28 October 2010 (has links)
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Previous issue date: 2010-10-28 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / In the present study, extracts rich-sulfated polysaccharides were obtained from three different species of Dictyotales (a class of brown macroalgae): Canistrocarpus cervicornis, Dictyota mertensii and Dictyopteris delicatula and their anticoagulant and antioxidant activities were evaluated. All extracts showed anticoagulant activity on aPTT assay, but not on PT assay. Extracts also exhibited total antioxidant activity, superoxide radical scavenging capacity and ferric chelating property. The extract from C. cervicornis showed the best results and was choose to have their sulfated polysaccharides fractioned and subsequently analysed. Thus, six fractions (CC-0.3, CC-0.5, CC-0.7, CC-1.0, CC-1.2 and CC-2.0) were obtained by proteolysis followed by sequential acetone precipitation. Agarose gel eletrophoresis stained with blue toluidine, confirmed the presence of sulfated polysaccharides in all fractions. Chemical analyses showed that all fractions presented heterofucans mainly constitued by fucose, galactose, glucuronic acid and sulfate. Any fraction changed the PT. However, all fractions were able to double the aPTT on a dose-dependent manner. CC- 0.3, CC-0.5, CC-0.7 and CC-1.0 needed only 0.100 mg/mL to double the aPTT, result only 1.25 times higher than the Clexane? (0.080 mg/mL), a commercial low molecular heparin. The heterofucans presented appreciable total antioxidant capacity, low capacity on scavenging hydroxyl radical and good efficiency on scavenging superoxide radicals (except CC-1.0). CC-1.2 showed 43.1 % on superoxide radical scavenging. This result was higher than that showed by the same concentration of gallic acid (41.8 %), a known antioxidant. Furthermore, the heterofucans showed excelent activity on ferrous chelating activity (except CC-0.3). CC-0.5, CC-0.7 and CC-1.0 showed the highest activities with 47.0 % of ferrous chelating activity, a result 2.0 times lesser than that exhibited by the same concentration of EDTA. These results clearly indicated the beneficial effects of heterofucans extracted from C. cervicornis as potential anticoagulant and antioxidant agents. However additional steps of purification, structural studies, besides in vivo experiments are needed for these fucans may be used as therapeutic agents / No presente estudo, extratos brutos ricos em polissacar?deos sulfatados foram obtidos a partir de tr?s esp?cies de Dictyotales (uma ordem de macroalgas marrons): Canistrocarpus cervicornis, Dictyota mertensii e Dictyopteris delicatula, e suas atividades anticoagulante e antioxidante foram avaliadas. Todos os extratos apresentaram atividade anticoagulante frente ao ensaio de aPTT, mas n?o sobre o ensaio de PT. Os extratos tamb?m exibiram atividade antioxidante total, capacidade em sequestrar radicais super?xido e propriedade de quelar ferro. O extrato obtido a partir de C. cervicornis apresentou os melhores resultados e foi escolhido para ter seus polissacar?deos sulfatados fracionados e subsequentemente analisados. Deste modo, seis fra??es (CC-0.3, CC-0.5, CC-0.7, CC-1.0, CC-1.2 e CC-2.0) foram obtidas por prote?lise seguida de precipita??o sequencial com acetona. Eletroforese em gel de agarose corada com azul de toluidina comprovou a presen?a de polissacar?deos sulfatados em todas as fra??es. As an?lises qu?micas mostraram que todas as fra??es apresentam heterofucanas constitu?das principalmente por fucose, galactose, ?cido glucur?nico e sulfato. Nenhuma fra??o alterou o PT. Entretanto todas as fra??es foram capazes de dobrar o aPTT de uma maneira dose dependente. As fra??es, CC-0.3, CC-0.5, CC-0.7 e CC-1.0, precisaram de apenas 0,100 mg/mL para dobrar o tempo de coagula??o, concentra??o que ? apenas 1,25 vezes maior do que aquela utilizada com Clexane? (heparina de baixo peso molecular) para se obter o mesmo efeito. As heterofucanas apresentaram apreci?vel capacidade antioxidante total, baixa capacidade em sequestrar radicais hidroxila e uma boa efici?ncia em sequestrar radicais super?xido (exceto CC-1.0). CC-1.2 mostrou uma capacidade de sequestrar 43,1 % dos radicais super?xido. Este resultado foi maior do que o apresentado pela mesma concentra??o de ?cido g?lico (41,8 %), um antioxidante conhecido. Al?m disso, as heterofucanas mostraram uma excelente atividade em quelar ferro (exceto CC-0.3). CC-0.5, CC-0.7 e CC-1.0 apresentaram as maiores atividades com 47,0 % de quela??o f?rrica, um resultado 2 vezes menor do que o exibido pela mesma concentra??o de EDTA. Estes resultados indicaram claramente os efeitos ben?ficos de heterofucanas extra?das de C. cervicornis como potenciais agentes anticoagulante e antioxidante. Entretanto, passos adicionais de purifica??o, estudos estruturais, al?m de experimentos in vivo, s?o necess?rios para que estas fucanas possam vir a ser utilizadas como agentes terap?uticos
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Avaliação da atividade vasorrelaxante da alga marinha brasileira Dictyota pulchella Hörning & Schnetter em ratos normotensos / Evaluation of vasorelaxant activity from brazilian marine algae Dictyota pulchella Hörnig & Schnetter in normotensive rats.Queiroz, Thyago Moreira de 21 February 2011 (has links)
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Previous issue date: 2011-02-21 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The pharmacological effects induced by CH2Cl2/MeOH extract (EDP) and
Hexane/EtOAc phase (FDP) from the Brazilian alga Dictyota pulchella were studied
on the cardiovascular system of Wistar rats using a combined in vivo and in vitro
approach. All protocols in this study were approved by the CEPA/LTF (protocol nº
0208/10). In normotensive conscious male rats, EDP injections (5; 10; 20 and 40
mg/kg, i.v., randomly) produced hypotension (-4.1 ± 1.34; -7.0 ± 2.4; -46.9 ± 1.3 and -
54.8 ± 4.3%; respectively) and bradycardia (-2.1 ± 1.6; -4.0 ± 2.3; -66.8 ± 5.2 and -
74.7 ± 4.5%; respectively) (n=5). Isolated superior mesenteric artery rings (1-2 mm)
were suspended by cotton threads for isometric tension recordings in a Tyrode s
solution at 37 ºC, gassed with a 95% O2 and 5% CO2, under a resting tension of
0.75g. In phenylephrine (Phe, 1μM)-pre-contracted rings, EDP (0.01 500 μg/mL)
induced a concentration-dependent relaxation (Maximum Response = 101.4 ± 4.5%;
EC50 = 22.35 ± 5.09 μg/mL) and this effect was not modified by removal of the
vascular endothelium (MR = 103.3 ± 8.3%; EC50 = 21.43 ± 8.98 μg/mL, n=7). Similar
results were found in the presence of FDP (0.01 500 μg/mL). FDP induced a
concentration-dependent vasodilatation in both endothelium-intact (MR = 80.6 ±
5.8%; EC50 = 24.1 ± 8.95 μg/mL, n=6) or endothelium-denuded mesenteric artery
rings (MR = 95.6 ± 7.5%; EC50 = 23.7 ± 5.65 μg/mL, n=6). Based on the preliminary
results, the subsequent experiments were performed in rings without endothelium. To
appreciate the involvement of potassium channels, the preparations were preincubated
with Tyrode s modified solution, KCl (20 mM) or with non-selective K+
channel blocker, tetraethylammonium (TEA, 3 mM). In both preparations the
vasorelaxant activity was not changed. In the presence of a tromboxane A2 agonist
U-46619 (100 nM), EDP induced concentration-dependent vasodilatation (MR = 90.3
± 7.8%; EC50 = 24.63 ± 4.04 μg/mL, n=6) was similar to the response found under
Phe-induced. After exposure to high concentrations of extracellular K+ (KCl, 60 mM),
the EDP induced concentration-dependent vasodilatation (MR = 97.7 ± 4.0%; EC50 =
34.57 ± 5.11 mg/mL; n=6). In the same experimental condition, FDP induced
concentration-dependent vasodilatation (MR = 113.5 ± 6.1%; EC50 = 10.92 ± 2.81
μg/mL; n=6). This result indicates that both EDP and FDP act on voltage-operated
calcium channel (Cav). Furthermore, EDP and FDP (0.03; 0.3; 10; 30 e 100 μg/mL)
antagonized CaCl2-induced contractions. The extract also induced vasodilatation in
the contraction evoked by L-type Ca2+ channel agonist (Bay K 8644, 200 nM) (MR =
113.3 ± 6.7%; EC50 = 19.45 ± 6.66 μg/mL, n=7). These results suggest that EDP
induces hypotension and bradycardia. Both EDP and FDP induce endotheliumindependent
vasodilatation that involves the inhibition of the Ca2+ influx through
blockade of Cav. / Os efeitos farmacológicos do extrato CH2Cl2:MeOH (EDP) e fase Hexano:AcOEt
(FDP) da alga marinha brasileira Dictyota pulchella foram estudados sobre o sistema
cardiovascular de ratos, utilizando uma abordagem in vivo e in vitro. Em ratos
normotensos não anestesiados, EDP (5; 10; 20 e 40 mg/kg, i.v., randomicamente)
promoveu hipotensão (-4,1 ± 1,34; -7,0 ± 2,4; -46,9 ± 1,3 e -54,8 ± 4,3%,
respectivamente) acompanhada de bradicardia (-2,1 ± 1,6; -4,0 ± 2,3; -66,8 ± 5,2 e -
74,7 ± 4,5%, respectivamente) (n=5). Em anéis de artéria mesentérica superior
isolada de rato pré-contraídos com Fenilefrina (FEN) 1 μM, EDP (0,01 500 μg/mL)
promoveu um efeito vasorrelaxante dependente de concentração na presença do
endotélio vascular (Emáx = 101,4 ± 4,5%; CE50 = 22,35 ± 5,09 μg/mL), e este efeito
não foi alterado após a remoção do endotélio (Emáx = 103,3 ± 8,3%; CE50 = 21,43 ±
8,98 μg/mL) (n=7). Resultados semelhantes foram obtidos na presença de FDP
(0,01 500 μg/mL), observando-se um vasorrelaxamento tanto na presença (Emáx =
80,6 ± 5,8%; CE50 = 24,1 ± 8,9 μg/mL), quanto na ausência do endotélio funcional
(Emáx = 95,6 ± 7,5%; CE50 = 23,70 ± 5,65 mg/mL). Para avaliar se o efeito de EDP
era dependente do tônus vascular, este extrato foi testado no tônus basal, na
presença ou ausência do endotélio, demonstrando que a resposta não foi alterada
em nenhuma das duas situações. Baseado nos resultados preliminares, os
experimentos subseqüentes foram realizados com endotélio desnudo. Para avaliar a
participação dos canais para potássio (K+), utilizou-se uma solução com 20 mM de
KCl ou tetraetilâmonio (TEA) 3 mM. Em ambas as preparações (Emáx = 102,3 ±
4,8%; CE50 = 25,40 ± 6,05 μg/mL) ou (Emáx = 111,2 ± 5,3%; CE50 = 16,70 ± 3,61
μg/mL) (n=7), respectivamente, a atividade vasorrelaxante de EDP não foi alterada.
Na presença de outro agente contracturante, U46619 (100 nM), EDP promoveu um
efeito vasorrelaxante (Emáx = 90,3 ± 7,8%; CE50 = 24,63 ± 4,04 μg/mL) de maneira
similar aos anéis pré-contraídos com FEN. Em experimentos contendo uma solução
despolarizante de 60 mM de KCl, EDP causou vasorrelaxamento dependente de
concentração (Emáx = 97,7 ± 4,0%; CE50 = 34,57 ± 5,11 mg/mL; n=6). Na mesma
condição experimental, FDP também promoveu um efeito vasorrelaxante (Emáx =
113,5 ± 6,1%; CE50 = 10,92 ± 2,81 μg/mL; n=6), não havendo diferença significante,
para os dois compostos, quando comparados aos anéis pré-contraídos com FEN.
Sugere-se que tanto EDP quanto FDP atuem sobre os canais para cálcio sensíveis a
voltagem (Cav). Além disso, EDP e FDP (0,03; 0,3; 10; 30 e 100 μg/mL)
antagonizaram as contrações induzidas por CaCl2. O extrato ainda produziu
vasorrelaxamento na presença de um agonista de canais para Ca2+ tipo-L (Bay K
8644; 200 nM) (Emáx = 113,3 ± 6,7% e CE50 = 19,45 ± 6,66 μg/mL, n=7). Esses
resultados sugerem que EDP produz hipotensão e bradicardia transientes, e tanto
EDP quanto FDP promovem vasorrelaxamento independente do endotélio vascular
por inibição do influxo de Ca2+, por meio do bloqueio dos Cav.
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Macroalgal dynamics on Caribbean coral forereefsRenken, Hendrik January 2008 (has links)
Tropical coral reefs are among the most diverse ecosystems of the world but facing increasing threats to their health. Over the last thirty years, many Caribbean coral reefs have undergone dramatic changes and experienced large losses in coral cover, due to direct and indirect anthropogenic disturbances. The results of which are reefs with low rugosity, changed trophic dynamics and low fish diversity. In recent times reefs have failed to recover from disturbances due to an increase in frequency and severity of disturbances and stresses. In the Caribbean on many coral reefs this has resulted in a shift towards macroalgal dominance by species of the phylum Phaeophyta. The processes and factors affecting the standing crop of macroalgae are many and complex. Two main hypotheses are identified in the literature as being the driving forces of algal dynamics: nutrient dynamics (availability, supply and uptake) and herbivory. However, many studies have been found to be inconclusive because of the complexity of the coral reef ecosystem, which makes it difficult if not impossible to control for all factors and processes influencing the standing crop of macroalgae such as light, water flow and sedimentation. The inherent characteristics of macroalgae, like morphology and life history, make them behave differently. Whilst herbivore characteristics, like size of mouth parts, feeding modes and preferences, will influence the amount of algal biomass removed. The spatial context (i.e. coral fore reef vs. back reef) will influence the effects of both bottom-up and top-down controls. Besides these inter-habitat differences, macroalgae within similar habitats but differing geographical locations may respond differently, for example, a forereef exposed to the open ocean or a forereef located in a sheltered bay. This thesis attempts to provide insight into the dynamics of two dominant brown macroalgae on Caribbean coral reefs, Dictyota spp. and Lobophora variegata. This aim was addressed by developing a model for the macroalga species Dictyota to model the various processes and factors on a coral forereef affecting percentage cover. Further, the patch dynamics of both Lobophora variegata and Dictyota were investigated to gain an insight into their dynamics under varying environmental conditions: the windward and leeward sides of an atoll. Finally, herbivory is identified as one of the key process affecting macroalgal cover. I investigated this process by deploying cages on both the windward and leeward side of the atoll to investigate the effects of grazing pressure under varying environmental conditions. A Bayesian Belief Network model was developed for Dictyota spp. to model the bottom-up and top-down processes on a coral forereef determining the percentage cover. The model was quantified using relationships identified in the scientific literature and from field data collected over a nine moth period in Belize. This is the first BBN model developed for brown macroalgae. The fully parameterized model identified areas of limited knowledge and because of its probabilistic nature it can explicitly communicate the uncertainties associated with the processes and interactions on standing crop. As such the model may be used as a framework for scientific research or monitoring programmes and it is expected that the model performance to predict macroalgal percentage cover will improve once new information becomes available. Size-based transition matrices were developed for both Dictyota spp. and Lobophora variegata to investigate the patch dynamics under varying environmental conditions: the windward and leeward sides of an atoll. The matrices reveal that standard measures of algal percent cover might provide a misleading insight into the underlying dynamics of the species. Modelling the patch dynamics with matrices provided insight into the temporal behaviour of macroalgae. This is an important process to understand because patch dynamics are determining competitive interactions with other coral reef benthic organisms. The outcome of competitive interactions will differ with macroalgal species. This study indicate that Dictyota spp. responded strongly to differing environmental conditions in that it has reduced growth rates and lower percent cover on the leeward side of the atoll, whilst Lobophora variegata showed far less sensitivity to environmental conditions. The patch dynamics of Dictyota spp. also showed a higher temporal variation than Lobophora variegata but only on the exposed forereef. A caging experiment was set up to investigate the response of both macroalgal species to different grazing pressure scenarios, under varying environmental conditions. Dictyota spp. had a significant response to environmental conditions in that a higher percentage cover was found on the exposed side of the atoll, whilst for Lobophora variegata the response was far less obvious. The less clear response of Lobophora variegata was very likely caused by competition of Dictyota with Lobophora due to the very high cover Dictyota obtained in the cages where all herbivores were excluded. The low grazing pressure treatments also showed an increase in cover of Dictyota, whilst for Lobophora, only a reduction in the rate of increase could be observed. The results indicate that on the leeward side of the atoll, fish grazing alone seems sufficient to control the standing crop of Dictyota and Lobophora variegata. Retrospective analysis of the experimental design showed that the limited size of the experimental set up could have confounded the results for Lobophora as well. In future experiments it is recommended to increase number replicates. Management of coral reef habitats is frequently constrained by a lack of funds and resources. The BBN Model once fully parameterized can provide a useful tool for coral reef management, because the model allows exploration of different reef scenario’s, which in turn can aid in prioritizing management strategies. Furthermore, the thesis provided an insight into the complexities of macroalgal dynamics. The responses of macroalgae to physiological factors and ecological processes are species specific and dependent on the location, and caution against generalizing on what controls the standing crop of macroalgae. Therefore it is argued that future investigations into algal ecology should clearly define the species, habitat and location. This can help to make informed management decisions.
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Recherche de molécules non-toxiques actives en antifouling à partir d'organismes marins de MéditerranéeViano, Yannick 16 June 2010 (has links) (PDF)
Alors que les premières formes de vie sont apparues dans les océans il y a près de 4 milliards d'années, les premières espèces terrestres remontent seulement à 400 millions d'années. Malgré cela, les substances naturelles d'origine marine ne représentent que 10% de l'ensemble des molécules isolées à ce jour à partir d'organismes vivants. Ces composés chimiques sont pourtant très spécifiques du fait des particularités du Monde Marin. Au sein des organismes qui les biosynthétisent, ils peuvent notamment intervenir dans la défense chimique contre les parasites et les autres compétiteurs. Ces molécules apparaissent donc comme des alternatives potentielles aux oxydes du tributylétain (TBTO) présents dans les revêtements antifouling et interdits depuis 2008 en raison de leur toxicité. Dans ce contexte et dans le cadre d'un partenariat avec le Parc national de Port-Cros, l'étude de la composition chimique de plusieurs organismes marins méditerranéens, des algues vertes (Caulerpa taxifolia) et brunes (Dictyota sp., Dictyota dichotoma, Cystoseira foeniculacea) ainsi que des bryozoaires, a été entreprise. Ces travaux ont permis l'isolement et la caractérisation structurale d'une vingtaine de métabolites secondaires (dérivés terpéniques et lipidiques, stérols), parmi lesquels huit sont originaux. L'identification de l'ensemble de ces molécules a été réalisée par le biais de l'utilisation de différentes méthodes spectroscopiques (RMN 1D et 2D, SM-HR) et la stéréochimie de certaines d'entre-elles a été établie par RMN (expérience NOESY 1H-1H) et modélisation moléculaire. Des corrections de données spectrales ont été également proposées pour cinq composés connus. Par ailleurs, l'activité anti-adhésion de plusieurs métabolites issus d'algues a été évaluée vis-à-vis d'un biofilm bactérien marin (Pseudoalteromonas sp.) afin de déterminer leur potentiel en tant qu'agents antifouling : plusieurs de ces composés présente une activité intéressante (CE50 = 30 μM), certes inférieure à celle du TBTO (CE50 = 10 μM), mais largement supérieure à celles de co-biocides utilisés actuellement. Parallèlement à ces travaux, l'analyse des variations temporelles et spatiales de l'expression métabolique de certaines des espèces étudiées a été réalisée à des fins écologiques. D'une part, les extraits mensuels de C. taxifolia présentent généralement une activité anti-adhésion importante quand le taux de caulerpényne dans l'extrait est élevé. D'autre part, d'importantes fluctuations ont notamment été observées dans la composition chimique des bryozoaires Myriapora truncata et Pentapora fascialis en fonction de leur lieu de récolte.
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Caracteriza??o estrutural e avalia??o das atividades farmacol?gicas da fucana B extra?da da alga Dictyota menstrualisCosta, Thiago Gomes 06 February 2014 (has links)
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Previous issue date: 2014-02-06 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Seaweeds are a major source of biologically active compounds . In the extracellular matrix of these organisms are sulfated polysaccharides that functions as structural components preventing it against dehydration. The fraction 0.9 (FucB) rich in sulfated fucans obtained from brown seaweed Dictyota menstrualis was chemical characterized and evaluated for pharmacological activity by testing anticoagulant activity, stimulatory action on the synthesis of an antithrombotic heparan sulfate, antioxidant activity and its effects in cell proliferation. The main components were FucB carbohydrates (49.80 ? 0.10 %) and sulfate (42.30 ? 0.015 %), with phenolic compounds ( 3.86 ? 0.016 %) and low protein contamination ( 0.58 ? 0.001 % ) . FucB showed polydisperse profile and analysis of signals in the infrared at 1262, 1074 and 930 cm -1 and 840 assigned to S = O bonds sulfate esters , CO bond presence of 3,6- anhydrogalactose , β -D- galactose non- sulfated sulfate and the axial position of fucose C4 , respectively. FucB exhibited moderate anticoagulant activity , the polysaccharides prolonged time (aPTT ) 200 ug ( > 90s ) partial thromboplastin FucB no effect on prothrombin time (PT), which corresponds to the extrinsic pathway of coagulation was observed. This stimulation promoted fraction of about 3.6 times the synthesis of heparan sulfate (HS) by endothelial cells of the rabbit aorta ( RAEC ) in culture compared with cells not treated with FucB . This has also been shown to compete for the binding site with heparin. The rich fraction sulfated fucans exhibited strong antioxidant activity assays on total antioxidant (109.7 and 89.5 % compared with BHT and ascorbic acid standards ) , reducing power ( 71 % compared to ascorbic acid ) and ferric chelation ( 71 , comparing with 5 % ascorbic acid). The fraction of algae showed cytostatic activity on the RAEC cells revealed that the increase of the synthesis of heparan sulfate is not related to proliferation. FucB showed antiproliferative action on cell lines modified as Hela and Hep G2 by MTT assay . These results suggest that FucB Dictyota menstrualis have anticoagulant , antithrombotic , antioxidant potential as well as a possible antitumor action, promoting the stimulation of the synthesis of antithrombotic HS by endothelial cells and is useful in the prevention of thrombosis, also due to its inhibitory action on species reactive oxygen ( ROS ) in some in vitro systems , being involved in promoting a hypercoagulable state / Algas marinhas s?o uma das principais fontes de compostos biologicamente ativos. Na matriz extracelular desses organismos existem os polissacar?deos sulfatados que funcionam como componente estrutural prevenindo-a contra desidrata??o. A fra??o 0,9 (FucB) rica em fucanas sulfatadas obtida da alga marrom Dictyota menstrualis foi caracterizada quimicamente e avaliada quanto a atividade farmacol?gica por meio de ensaios de atividade anticoagulante, a??o estimulat?ria sobre a s?ntese de heparam sulfato antitromb?tico, atividade antioxidante e seus efeitos na prolifera??o celular. Os principais componentes da FucB foram carboidratos (49,80 ? 0,10%) e sulfato (42,30 ? 0,015%), apresentando compostos fen?licos (3,86 ? 0,016%) e baixa contamina??o prot?ica (0,58 ? 0,001%). FucB mostrou perfil polidisperso e sinais na an?lise de infravermelho em 1262, 1074 e 930 e 840 cm-1 atribu?dos a liga??es S=O de ?steres de sulfato, presen?a de liga??o C-O de 3,6-anidrogalactose, β-D-galactose n?o sulfatada e sulfato na posi??o axial do C4 da fucose, respectivamente. FucB exibiu moderada atividade anticoagulante, este polissacar?deo prolongou o tempo de tromboplastina parcial activada (aPTT) a 200 ug (>90s) n?o foi observado qualquer efeito de FucB sobre o tempo de protrombina (PT), que corresponde a via extr?nseca da coagula??o. Esta fra??o promoveu estimula??o cerca de 3,6 vezes na s?ntese de heparam sulfato (HS) pelas c?lulas endoteliais da aorta de coelho (RAEC), em cultura, quando comparadas com as c?lulas n?o tratadas com FucB. Esta tamb?m demonstrou competir pelo s?tio de liga??o com a heparina. A fra??o rica em fucanas sulfatadas exibiu forte a??o antioxidante sobre os ensaios de antioxidante total (109,7 e 89,5% comparados com padr?es BHT e ?cido asc?rbico), poder redutor (71% comparado ao ?cido asc?rbico) e quela??o f?rrica (71,5% comparando com ?cido asc?rbico). A fra??o dessa alga mostrou atividade citost?tica sobre as c?lulas RAEC revelando que o aumento da s?ntese de heparan sulfato n?o est? relacionado ? prolifera??o. FucB apresentou a??o antiproliferativa sobre linhagens celulares modificadas como Hela e Hep G2 pelo ensaio de MTT. Esses resultados sugerem que FucB de Dictyota menstrualis tem potencial anticoagulante, antitromb?tico, antioxidante bem como uma poss?vel a??o antitumoral, promovendo a estimula??o da s?ntese de HS antitromb?tico pelas c?lulas endoteliais, sendo ?til na preven??o da trombose, devido tamb?m a sua a??o inibit?ria sobre as esp?cies reativas do oxig?nio (ROS) em alguns sistemas in vitro, estando envolvidos na promo??o de estado de hipercoagulabilidade
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Coral recovery on phase-shifted reefs depend upon the type of macroalgae presentVoss, Justin N 07 December 2016 (has links)
The Florida Keys experienced some of the most drastic transitions from coral to macroalgae dominated states, known as phase-or regime-shifts, of any reefs in the Caribbean. Macroalgae on coral reefs lower coral recruitment by deterring coral settlement either directly through competition or indirectly by changing the chemical environment near the benthos. With evidence of species-specific interactions to coral-macroalgae competition, the type of macroalgae on a phase-shifted coral reef might be more important than just identifying a reef transition. To answer this question, I tested the effect of Laurencia intricata (a macroalgae related to the settlement inducing crustose coralline algae) and Dictyotaceae (known for its toxic or allelopathic compounds) on Porites astreoides planulae behavior, settlement and choice settlement preference, and post-settlement survival. I found that P. astreoides planulae show a positive response to chemical cues released from L. intricata, crustose coralline algae, and species in the Dictyotaceae family. However, the positive chemical cue response becomes algal-specific as larvae start probing for settlement substrate. Providing P. astreoides larvae with a choice between settlement substrates, revealed that the algal structure caused higher settlement next to L. intricata, while Dictyotaceae deterred larval settlement. It may be beneficial for larvae to settle next to L. intricata over Dictyotaceae algae. I identified that post-settlement survival was enhanced when P. astreoides larvae settled next to L. intricata while Dictyotaceae species did not enhance or deter post-settlement survival. These results indicate that coral larvae may be responding differently to a variety of chemical cues. Any chemical or physical cue from a reef may be used by coral larvae to identify and locate settlement substrate on a reef. Once they identify a reef’s location, they express a more selective behavior during settlement by avoiding Dictyotaceae macroalgae and favoring L. intricata. This suggests that the composition of a phase-shifted reef matters to coral recovery, not only that it has shifted to a dominated macroalgal state.
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