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Atividade antiviral de extratos brutos, ricos em polissacar?deos sulfatados, obtidos de macroalgas marrons e verdes contra o v?rus dengue 2Bezerra, Fabiana Lima 14 June 2016 (has links)
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Previous issue date: 2016-06-14 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq) / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / A dengue ? a mais importante arbovirose que afeta o homem e constitui-se em
um grave problema de sa?de p?blica. A doen?a ? end?mica em regi?es
tropicais e subtropicais, abrangendo mais de 100 pa?ses, correspondendo ?
metade da popula??o mundial. As epidemias s?o recorrentes e estima-se que
os v?rus da dengue causem 390 milh?es de infec??es a cada ano, sendo uma
importante causa de morbidade. Apesar desse cen?rio, atualmente, ainda n?o
se disp?e de um antiviral contra a dengue. Neste contexto, diversos estudos
relatam a atividade antiviral dos polissacar?deos sulfatados obtidos das algas
marinhas contra v?rus envelopados, cuja a??o est? associada ? interfer?ncia
nas etapas iniciais do processo de infec??o viral. Neste estudo, avaliou-se a
potencial atividade antiviral de extratos brutos ricos em polissacar?deos
sulfatados obtidos das algas marrons Dictyota menstrualis (EBDMens) e
Dictyota mertensii (EBDM) e das algas verdes Codium isthmocladum (EBCI) e
Caulerpa sertularioides (EBCS) contra o v?rus dengue 2 (DENV-2), em
linhagem de c?lulas Vero, usando diferentes estrat?gias metodol?gicas
(tratamento simult?neo, tratamento p?s-infec??o, pr?-tratamento da c?lula, pr?tratamento
do v?rus, adsor??o, p?s-adsor??o e penetra??o). A citotoxicidade
dos extratos foi analisada pelo ensaio de redu??o do MTT. O estudo da
atividade antiviral foi determinado pela quantifica??o do n?mero de c?pias do
RNA viral por RT-qPCR ap?s 120 h de infec??o. Nenhum extrato exibiu
toxicidade para as c?lulas Vero no ensaio de redu??o do MTT na concentra??o
de 100 ?g/mL. Todos os extratos exibiram atividade antiviral quando
adicionados durante os primeiros 90 minutos de infec??o, demonstrando
redu??o significativa no n?mero de c?pias do RNA viral ap?s 120 horas. Os
extratos EBDMens e EBDM foram mais eficazes nos ensaios de pr?-tratamento
da c?lula e do v?rus, e o primeiro exerceu atividade antiviral superior ao EBDM
durante a adsor??o viral. Os extratos EBCI e EBCS apresentaram atividade
antiviral semelhante no ensaio de pr?-tratamento da c?lula. O EBCI foi mais
eficaz na redu??o da adsor??o do DENV-2 em c?lulas Vero. Por?m, o EBCS se
mostrou mais eficiente nos ensaios de pr?-tratamento do v?rus e penetra??o.
Dentre os extratos avaliados, o EBCS parece ser o mais efetivo no combate ao
DENV-2. Estes resultados revelam o potencial desses extratos ricos em
polissacar?deos sulfatados como compostos com a??o antiviral, sugerindo que
eles atuam nos est?gios iniciais da infec??o por DENV-2. / Dengue is considered the most important human arboviruses, and is a serious
public health problem. This disease is endemic in tropical and subtropical
regions, reaching more than 100 countries, which means the half of world
population, and those epidemies have been appellant. It is estimated that
Dengue virus could cause 390 millions of infections each year, and is an
important cause of morbidity. Although this scenario, actually doesn?t exist a
dengue antivirals still. Sorts of studies have showed antivirus activity of
seaweed sulfated polysaccharides against enveloped viruses, which action
seems to be associated to initials steps of infection process. In this study, was
evaluated the potential antiviral activity of crude extracts rich in sulfated
polysaccharides obtained from the brown algae Dyctiota menstrualis
(EBDMens) and Dyctiota mertensii (EBDM), and green algae Codium
isthmocladium (EBCl) and Caulerpa sertularioides (EBCS), tested against
Dengue virus 2 (DENV-2) in Vero cell line, by using different methodological
strategies (simultaneous treatment, post-infection treatment, cell pre-treatment,
virus pre-treatment, adsorption, post-adsorption and penetration). The extracts
cytotoxicity was evaluated by MTT reduction assay. The study of antiviral
activity was determinated by quantifying viral RNA load using RT-qPCR, after
120 hours of infection. None extract has showed toxicity against Vero Cells in
MTT reduction assay with 100 ?g/mL concentration. All extracts have shown
antiviral activity when added during the first 90 minutes of infection, and a
significant reduction of viral RNA number of copies after 120 hours. The
EBDMens and EBDM extracts were more efficient to cell and virus pretreatment
assays, and the first has showed higher antiviral activity than EBDM
during viral adsorption. The EBCI and EBCS extracts have presented antiviral
activity very similar in cell pre-treatment assay. EBCI was more efficient in the
reduction of DENV-2 adsorption in Vero cell. However, EBCS has showed more
efficient in cell pre-treatment and penetration assays. Between the extracts
evaluated, EBCS seems to be more effective against DENV-2. These results
have showed potential action of extracts rich in sulfated polysaccharides with
antivirus activity, suggesting that they act in the initials steps of infection
process of DENV-2.
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S?ntese verde de nanopart?culas contendo prata e uma fra??o da Alga Spatoglossum schr?ederi composta por ?cido alg?nico e fucana A: caracteriza??o f?sico-qu?mica e avalia??o da atividade antiproliferativa frente ?s c?lulas de melanoma (B16F10)Dantas, Larisse Ara?jo 17 April 2017 (has links)
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Previous issue date: 2017-04-17 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq) / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / Nanopart?culas de prata (AgNPs) possuem diversas aplica??es biom?dicas, dentre elas destaca-se o potencial antiproliferativo. In?meros tipos de s?ntese de nanopart?culas de prata encontram-se descritos na literatura, no entanto, um tipo de s?ntese menos agressiva e mais economicamente vi?vel, a s?ntese verde, tem ganhado import?ncia dentre as demais. Neste processo de s?ntese ? necess?rio o uso de um agente redutor e estabilizante que n?o sejam t?xicos para as c?lulas animais e para o meio ambiente. Nesse contexto, a associa??o de biomol?culas de origem marinha com atividades biol?gicas j? descritas, como os polissacar?deos ?cidos de algas, ?s AgNPs ganham import?ncia. Muitos polissacar?deos ?cidos (PA) extra?dos de algas marinhas destacam-se por exibir diversas atividades, como exemplo as fucanas da alga Spatoglossum schr?ederi. Estas fucanas apresentam atividade antioxidante, antiangiog?nica, antitumoral, dentre outras. Devido aos poucos relatos de s?ntese de AgNPs envolvendo polissacar?deos de algas, este trabalho teve por objetivo sintetizar AgNPs com uma fra??o rica em ?cido alg?nico e fucana A da alga S. schr?ederi por um processo de s?ntese verde, e testar as AgNPs obtidas frente ? linhagem de melanoma B16F10. Os PA da alga S. schr?ederi foram extra?dos por um processo de digest?o proteol?tica e fracionados com acetona. A fra??o de maior rendimento e rica em ?cido alg?nico e fucana A, a F0.5 v, foi empregada para a s?ntese das AgNPs. A caracteriza??o das AgNPs obtidas contemplou a an?lise de absor??o de luz UV, composi??o qu?mica, di?metro m?dio, polidispers?o, potencial zeta, espectroscopia de infravermelho (FTIR) e a espectroscopia de energia dispersiva (EDS), microscopia eletr?nica de varredura (MEV) e microscopia de for?a at?mica (MFA). As an?lises por espectroscopia de UV-vis?vel na faixa entre 400 e 440 nm confirmaram a forma??o das AgNPs, estas apresentaram di?metro de 196 ? 13 nm, valores de polidispers?o abaixo de 0,4 e potencial zeta negativo. As dosagens qu?micas, o FTIR e o EDS revelaram que a maior parte das AgNPs ? composta por polissacar?deos ?cidos. As imagens obtidas por MEV e MFA indicaram um formato esf?rico para as AgNPs. A atividade antiproliferativa foi avaliada pelo m?todo do MTT frente ?s linhagens celulares B16F10 e fibroblastos 3T3, e por citometria de fluxo foi investigado sua a??o na morte e no ciclo celular. As AgNPs afetaram a capacidade de redu??o do MTT das linhagens 3T3 e B16F10, sendo esta mais evidenciada para a B16F10, pois as AgNPs (0,5 mg/mL) conseguiram reduz?-la em torno de 50%, enquanto a prata i?nica e a F0.5 v em nenhuma das concentra??es influenciaram na redu??o do MTT. As an?lises por citometria de fluxo apontaram uma elevada taxa de marca??o para anexina V e iodeto de prop?dio para as c?lulas B16F10 expostas ?s AgNPs, al?m de um aumento na porcentagem das c?lulas em Sub-G1 em detrimento ?s outras fases. Estudos adicionais s?o necess?rios para elucidar completamente o mecanismo de a??o antiproliferativo das AgNPs e descobrir outras propriedades da fra??o que s?o intensificadas pela s?ntese de nanopart?culas. Entretanto, pode-se concluir que ? poss?vel sintetizar AgNPs com polissacar?deos ?cidos de S. schr?ederi usando um m?todo verde e que essas AgNPs apresentaram atividade antiproliferativa mais pronunciada que seus percussores. / Silver nanoparticles (AgNPs) have several biomedical applications, among them the antiproliferative potential. Numerous types of synthesis of silver nanoparticles are found in the literature, however, a type of synthesis less aggressive and more economically viable, a green synthesisl, has gained value among the others. In this synthesis process, it is necessary to use a reducing agent and stabilizer which are non-toxic to animal cells and to the environment. In this context, an association of biomolecules of marine origin with biological activities already described, such as the polysaccharide acids of algae, to the AgNPs gain importance. Many acidic polysaccharides (PA) extracted from marine algae are distinguished by their different activities, such as the algae Spatoglossum schr?ederi. These fucans have antioxidant, antiangiogenic, antitumor activity, among others. Due to the few reports of AgNPs synthesis involving algae polysaccharides, the objective of this work was to synthesize AgNPs with a rich fraction of alginic acid and fucan A from the S. schr?ederi algae by a green synthesis process and to test the AgNPs obtained on the lineage of melanoma B16F10. The PA of the S. schr?ederi algae was extracted by a proteolytic digestion process and fractionated with acetone. The higher yield fraction rich in alginic acid and fucan A, F0.5 v, was used for synthesis of the AgNPs. The characterization of the AgNPs obtained contemplated an analysis of UV light absorption, chemical composition, average diameter, polydispersity, zeta potential, infrared spectroscopy (FTIR), dispersive energy spectroscopy (EDS), scanning electron microscopy (SEM) and atomic force microscopy (AFM). The analyzes by UV-visible spectroscopy in the range between 400 and 440 nm confirmed the formation of AgNPs, they presented 196 ? 13 nm diameter, polydispersion values below 0.4 and negative zeta potential. The chemical dosages, FTIR and EDS revealed that most AgNPs are composed of acidic polysaccharides. The images obtained by SEM and AFM indicated a spherical format for AgNPs. The antiproliferative activity was evaluated by the MTT method against B16F10 cell lines and 3T3 fibroblasts, and by flow cytometry, its action on death and cell cycle was investigated. The AgNPs affected the ability to reduce MTT from the 3T3 and B16F10 lineages, but these were evidenced for a B16F10, the AgNPs (0.5 mg/mL) were able to reduce by around 50%, while a silver ion and F0.5 v at any of the concentrations influenced the reduction of MTT. Flow cytometry analyzes indicated a high labeling rate for annexin V and propidium iodide for the B16F10 cells exposed to AgNPs, in addition to an increase in the percentage of cells in SubG1 in detriment to the other phases. Additional studies are needed to fully elucidate the mechanism of antiproliferative action of AgNPs and to discover other properties of the fraction that are enhanced by the synthesis of nanoparticles. However, it can be concluded that it is possible to synthesize AgNPs with acidic polysaccharides from S. schr?ederi using a green method and that these AgNPs showed more pronounced antiproliferative activity than their precursors.
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Caracteriza??o do efeito antinociceptivo e anti-inflamat?rio do polissacar?deo extra?do da levedura Kluyveromyces marxianusOliveira, Renata Freitas de Araujo 26 May 2015 (has links)
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Previous issue date: 2015-05-26 / The yeasts produce different types of metabolites and macromolecules with huge biotechnological potential. Among them, the Kluyveromyces marxianus, species attracts industrial and economic interests for presenting certain qualities, such as thermal tolerance, high growth rates, broad substrate spectrum and modulation of immune response. This work investigated the effects of treatment with the polysaccharide extracted from the Kluyveromyces marxianus yeast (poly322) in pain and inflammation experimental models. The poly322 pharmacological properties were evaluated with writhing, formalin, nociception (manifested by CFA), tail flick, hot-plate and paw edema tests. The cytokine levels were determined by ELISA and the effects on motor performance were evaluated by rotarod and open field tests. The pretreatment with poly322 resulted in reduced nociception induced by acetic acid, CFA and formalin (late phase). Furthermore, animals treated with poly322 exhibited a significant reduction in mechanical hypernociception, paw edema, and local IL-6 levels increase induced by carrageenan. In contrast, treatment with poly322 did not alter the thermal stimulus response threshold in the tail flick and hot plate tests, indicating no central action. Confirming the specificity of the action suggested by nociceptive tests, the treatment with poly322 did not induce motor impairment. These results demonstrate that the poly322 has a potent antinociceptive and anti-inflammatory effects, possibly mediated by the inhibition of the production and/or release of the IL-6 cytokine. / As leveduras produzem diferentes tipos de metab?litos e macromol?culas com enorme potencial biotecnol?gico. Dentre essas, destaca-se a Kluyveromyces marxianus, esp?cie que desperta interesse industrial e econ?mico por apresentar qualidades, como a toler?ncia t?rmica, altas taxas de crescimento, amplo espectro de substrato e modula??o da resposta imune. O presente estudo investigou os efeitos do tratamento com o polissacar?deo extra?do da levedura Kluyveromyces marxianus (poly322) em modelos experimentais de dor e inflama??o. As propriedades farmacol?gicas do poly322 foram avaliadas nos testes de contor??es abdominais, formalina, nocicep??o manifesta por CFA, retirada da cauda, placa quente e edema de pata. Os n?veis de citocinas foram determinados por ELISA e os efeitos sobre o desempenho motor foram avaliados pelos testes do cil?ndro girat?rio e campo aberto. O pr?-tratamento com o poly322 resultou na redu??o da hipernocicep??o induzida pelo ?cido ac?tico, CFA e formalina (segunda fase). Al?m disso, os animais tratados com o poly322 exibiram uma redu??o significativa no edema de pata e no aumento local dos n?veis de IL-6 induzidos pela carregenina. Em contraste, o tratamento com o poly322 n?o alterou o limiar de resposta a est?mulo t?rmico nos testes de retirada da cauda e placa quente, indicando aus?ncia de a??o central. Confirmando a especificidade da a??o sugerida pelos testes nociceptivos, o tratamento com o poly322 n?o induziu comprometimento motor. Os resultados demonstram que o poly322 possui um potente efeito antinociceptivo e anti-inflamat?rio, possivelmente mediado pela inibi??o da produ??o e/ou libera??o da citocina IL-6.
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Bioprospec??o qu?mica da esponja callyspongia vaginalis (Callyspongiidae)Ara?jo, Rusceli Diego de 05 April 2013 (has links)
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Previous issue date: 2013-04-05 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Recently, marine organisms have attracted attention because of the
complexity and potent biological activity from your secondary metabolites. Our
planet has 80% it surface covered by oceans and seas, therefore, housing a
wide number of different forms of life, among them, the sponges. These sessile
and filtrating animals, according to numerous researches, come showing like
true chemistry factories. The substances from these animals, sometimes show
as news targets to therapeutics agents, and some countries has already use
them for treatment of some diseases. Further of the secondary metabolites, the
polysaccharides of marine origin also have been target of studies, because the
presence of the sulfates groups in its molecules. Polysaccharides with differents
biological activities have been related in a large number of researches. Actually,
many studies show the sponges as source of promising medicine. These
studies inspire new researches, because the few number of sponges species
studied until now. Because of that, the present work shows the chemistry
prospection of the sponge Callyspongia vaginalis. Chromatographic methods in
silica gel allowed the isolations of two secondary metabolites: the known β-
sitosterol and a ceramide, no reported in the genus Callyspongia, previously.
The analysis of the their lipid extracts show different kinds of fatty acids with a
variety of chain length (saponifiable fraction), and others metabolites like
Lupenone and stigmasterol, also unprecedented in the genus. The
Polysaccharide characterization and the elucidation of the secondary
metabolites acquired through of chromatography analysis (CC, molecular
exclusion) and spectrometric (NMR 1H and 13C, mass, IR), respectively and
comparison with literature data / A diversidade estrutural de subst?ncias de origem marinha, associadas
as suas atividades biol?gicas, t?m atra?do aten??o de pesquisadores do mundo
inteiro. Dentre os organismos estudados atualmente dois grupos de
invertebrados marinhos, as asc?dias e as esponjas, merecem destaque. Estes
animais s?sseis e filtradores, segundo in?meras pesquisas, v?m se mostrando
verdadeiras f?bricas qu?micas , produzindo subst?ncias que por vezes
apresentam-se como aspirantes a agentes terap?uticos, com efic?cia cl?nica
comprovada. Al?m dos metab?litos secund?rios, os polissacar?deos de origem
marinha tamb?m tem sido alvo de estudos, sendo relatados diversos
compostos sulfatados e com diferentes atividades biol?gicas associadas a
estes. Tendo em vista a pequena quantidade de esp?cies de esponjas
estudadas at? o momento, comparado ao potencial qu?mico destes
organismos, o presente trabalho apresenta a prospec??o qu?mica da esponja
Callyspongia vaginalis. T?cnicas cl?ssicas de cromatografia em gel de s?lica
permitiram o isolamento de dois metab?litos secund?rios: o β-sitosterol e uma
ceramida, ambos in?ditos no g?nero Callyspongia. A an?lise das fra??es
lip?dicas mostraram diferentes tipos de ?cidos graxos com variados tamanhos
de cadeia (fra??o saponific?vel), e de outros metab?litos como lupenona e
estigmasterol, tamb?m in?ditos no g?nero em quest?o. O polissacar?deo obtido
atrav?s do material animal tamb?m foi isolado e, como amplamente relatado na
literatura apresentou-se sulfatado, caracter?stica diferencial dos polissacar?deos
de origem marinha. A caracteriza??o do polissacar?deos e a elucida??o
estrutural dos metab?litos secund?rios foram realizadas atrav?s da an?lise dos
dados de espectrometria de massa, Infravermelho e Resson?ncia Magn?tica
Nuclear de Hidrog?nio-1 e Carbono-13, atrav?s de sequ?ncias de pulsos uni e
bidimensionais e compara??o com dados da literatura
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Atividade antioxidante de fucanas e galactanas extra?das de algas marinhasSouza, Micheline Cristiane Rocha de 26 May 2008 (has links)
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Previous issue date: 2008-05-26 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Studies made with polysaccharides of seaweed have demonstrated that these present important biological and pharmacological activities. These composites had
presented "scavenging" activity of free radicals, which is important in the mediation of the inflammatory process and in the pathology of diverse disease. Recently, this "scavenging" property has taken some researches to evaluate the antioxidant
capacity from various polysaccharides. Considering the limited research with polysaccharides and knowing its largely employed by the pharmaceutical and foodstuffs industries, we have objective to verify the actions from fucans and galactans as antioxidants. The fucans are found in brown seaweed and the galactans (carrageenans) in red seaweed. The fucans were obtained from seaweed Padina gymnospora (F0.5 e F1.1 fractions), common to our coastline and one another fucan,
fucoidan, was of origin commercial and extracted from seaweed Fucus vesiculosus. The λ, κ e ι carrageenans were also of origin commercial. The antioxidant activities were tested in superoxide and hydroxyl systems to generated free radicals and for
the inhibition of the lipid peroxidation. The results obtained to inhibition of formation the superoxide radicals demonstrated that all polysaccharides presented scavenging activity of superoxide radicals. The fucoidan, F0.5 and F1.1 fractions
presented IC50 of 0.058; 0.243 and 0.243 mg/mL, respectively, while IC50 of the λ, κ and ι carrageenans were 0.046; 0.112 and 0.332 mg/mL, respectively. The results to inhibition of formation the hydroxyl radicals demonstrated that all sample had low effect in the inhibition of the formation of these radicals, except the F0.5. For these radicals the IC50 were 0.157 and 0.353 mg/mL to the fucoidan and F1.1, respectively
and 0.357; 0.335 and 0.281 mg/mL to λ, κ and ι carrageenans, respectively. All the samples were capacity to inhibition the peroxidation, it present the IC50 of 1.250; 2.753 and 2.341 mg/mL to fucoidan, F0.5 and F1.1, respectively. Already the λ, κ and ι carrageenans presented the IC50 of 2.697; 0.323 and 0.830 mg/mL, respectively. With these findings, we conclude that polysaccharides used in this study presented activity antioxidant, and that fucoidan and the λ carrageenan show a significant "scavenging" activity for the radicals superoxide and the κ carrageenan a significant inhibitory activity for the lipid peroxidation / Estudos feitos com polissacar?deos de algas t?m demonstrado que esses apresentam atividades biol?gicas e farmacol?gicas importantes. Estes compostos apresentaram atividade varredora de radicais livres, os quais s?o importantes na
media??o do processo inflamat?rio e na patologia de diversas doen?as. Recentemente, essa propriedade varredora tem levado alguns pesquisadores a avaliar a capacidade antioxidante de diversos polissacar?deos. Considerando o reduzido n?mero de pesquisas com estes compostos e sabendo-se de seu largo
emprego pela ind?stria farmac?utica e aliment?cia, objetivamos neste trabalho, verificar as a??es de fucanas e galactanas como antioxidantes. As fucanas s?o encontradas em algas marrons e as galactanas (carragenanas) em algas vermelhas.
As fucanas foram obtidas da alga Padina gymnospora (fra??es F0,5 e F1,1), comum em nosso litoral, e uma outra fucana, o fucoidan, foi de origem comercial e extra?do da alga Fucus vesiculosus. As carragenanas λ, κ e ι tamb?m foram comerciais. A atividade antioxidante foi testada em sistemas geradores de radicais super?xidos e hidroxilas, e pela inibi??o da peroxida??o lip?dica. Os resultados obtidos para inibi??o da forma??o de radicais super?xidos demonstraram que todos os
polissacar?deos apresentaram atividade varredora de radicais super?xidos. O fucoidan, as fra??es F0,5 e F1,1 apresentaram IC50 de 0,058; 0,243 e 0,243 mg/mL, respectivamente; enquanto o IC50 das carragenanas λ, κ e ι foi de 0,046; 0,112 e 0,332 mg/mL, respectivamente. Os resultados para inibi??o da forma??o de radicais hidroxilas demonstraram que todas as amostras inibiram a forma??o destes radicais,
exceto a F0,5. Para estes radicais o IC50 foi de 0,157 e 0,353 mg/mL para o fucoidan e F1,1, e de 0,357; 0,335 e 0,281 mg/mL para carragenanas λ, κ e ι, respectivamente. Todas as amostras foram capazes de inibir a peroxida??o lip?dica com IC50 de 1,250, 2,753 e 2,341 mg/mL para o fucoidan, F0,5 e F1,1, respectivamente. J? as carragenanas λ, κ e ι apresentaram o IC50 de 2,697, 0,323 e 0,830 mg/mL, respectivamente. Com estes achados, conclu?mos que os
polissacar?deos usados neste estudo apresentam atividade antioxidante, e que o fucoidan e a carragenana λ exibem uma significante atividade varredora para os radicais super?xido e a carragenana κ uma significante atividade inibit?ria da
peroxida??o lip?dica
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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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Atividades antinociceptiva e anti-inflamat?ria de uma fra??o rica em heterogalactana sulfatada extra?da da alga Codium isthmocladum (Vickers 1905)Cordeiro, Sara Lima 11 March 2013 (has links)
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Previous issue date: 2013-03-11 / Sulfated polysaccharides comprise a complex group of macromolecules with a
range of several biological activities, including antiviral activity, anticoagulant,
antiproliferative, antiherp?tica, antitumor, anti-inflammatory and antioxidant. These
anionic polymers are widely distributed in tissues of vertebrates, invertebrates and
algae. Seaweeds are the most abundant sources of sulfated polysaccharides in
nature. The green algal sulfated polysaccharides are homo or heteropolysaccharides
comprised of galactose, glucose, arabinose and/or glucuronic acid. They are
described as anticoagulant, anti-inflammatory, antiviral, anti-angiogenic, antitumor
compounds. However, there are few studies about elucidation and evaluation of
biological/pharmacological effects of sulfated polysaccharides obtained from green
algae, for example, there is only one paper reporting the antinociceptive activity of
sulfated polysaccharides of these algae. Therefore this study aimed to obtain sulfated
polysaccharides of green seaweed Codium isthmocladum and evaluates them as
potential antinociceptive agents. Thus, in this study, the total extract of
polysaccharides of green alga C. isthmocladum was obtained by proteolytic
digestion, followed by fractionation resulting in five fractions (F0.3, F0.5, F0.7, F0.9
and F1.2) by sequential precipitation with acetone. Using the test of abdominal
contractions we observed that the fraction F0.9 was the most potent antinociceptive
aompound. F0.9 consists mainly of a sulfated heterogalactana. More specific tests
showed that Fo.9 effect is dose and time dependent, reaching a maximum at 90 after
administration (10 mg / kg of animal). F0.9 is associated with TRPV1 and TRPA1
receptors and inhibits painful sensation in animals. Furthermore, F0.9 inhibits the
migration of lymphocytes induced peritonitis test. On the other hand, stimulates the
release of NO and TNF-?. These results suggest that F0.9 has the potential to be
used as a source of sulfated galactan antinociceptive and anti-inflammatory / Os polissacar?deos sulfatados comp?em um grupo complexo de
macromol?culas com uma gama de importantes atividades biol?gicas, incluindo
atividade antiviral, anticoagulante, antiproliferativa, antiherp?tica, antitumoral, antiinflamat?ria
e antioxidante. Esses pol?meros ani?nicos s?o amplamente distribu?dos
em tecidos de vertebrados, invertebrados e algas. As algas marinhas s?o as fontes
mais abundantes de polissacar?deos sulfatados na natureza. Os polissacar?deos
sulfatados de algas verdes s?o homo ou heteropolissacar?deos compostos por
galactose, glicose, arabinose e/ou ?cido glucur?nico. Para estes pol?meros s?o
descritas atividades como anticoagulante, anti-inflamat?ria, antiviral,
antiangiog?nica, antitumoral. Por?m, ainda h? poucos estudos de elucida??o e
avalia??o de atividades biol?gicas/farmacol?gicas de polissacar?deos sulfatados
obtidos de algas verdes, por exemplo, h? somente um trabalho relatando a atividade
antinociceptiva de polissacar?deos sulfatados destas algas. Portanto este trabalho
teve como objetivo obter polissacar?deos sulfatados da alga verde Codium
isthmocladum e avalia-los como poss?veis agentes antinociceptivos. Assim, no
presente estudo, o extrato total de polissacar?deos da alga verde C. isthmocladum
foi obtido atrav?s de digest?o proteol?tica, seguida de fracionamento resultando em
cinco fra??es (F0.3; F0.5; F0.7; F0.9 e F1.2) por precipita??o sequencial com
acetona. Com o teste de contra??es abdominais observou-se que a fra??o F0.9 foi o
mais potente antinociceptivo. F0.9 ? composta principalmente por uma
heterogalactana sulfatada. Ensaios mais espec?ficos mostraram que o seu efeito ?
dose e tempo dependente, chegando ao m?ximo com 90 ap?s a administra??o (10
mg/kg de animal). F0.9 se associa a receptores TRPV1 e TRPA1 e inibe sensa??o
dolorosa em animais. Al?m disso, F0.9 inibe a migra??o de linf?citos em ensaios de
peritonite induzida. Por outro lado estimula a libera??o de NO e TNF-?. Esses
resultados sugerem que F0.9 tem potencial para ser usada como fonte de galactana
sulfatada antinociceptiva e anti-inflamat?ria
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Atividade anticoagulante, antioxidante e antitumoral de heterofucanas da alga Dictyopteris delicatula (Lamouroux, 1809)Magalhaes, Kaline Dantas 02 December 2011 (has links)
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Previous issue date: 2011-12-02 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / In the present study, six families of sulfated polysaccharides were obtained from seaweed Dictyopteris delicatula (Lamouroux, 1809) and their anticoagulant, antioxidant and antitumor activities were evaluated. All fractions showed anticoagulant activity on aPTT assay, but not on PT assay. Fractions also exhibited total antioxidant activity, superoxide radical scavenging capacity and ferric chelating property. Thus, six fractions (F0.5v, F0.7v, F1.0v, F1.3v, F1.5v e F2.0v) we obtained by proteolytic digestion, followed by acetone fractionation and molecular sieving on Sephadex G-100. Chemical analyses demonstrated that all polysaccharides contain heterofucans composed mainly of fucose, xylose, glucose, galactose, uronic acid, and sulfate. Any fractions changed the PT. However, all fractions were able on double the aPPT on a dose-dependent manner. The heterofucans F0.7v and F1.0v showed low anticoagulant activity while F1.5v presented the most prominent anticoagulant activity .When compared to Clexane?, a low molecular weight heparin, at same concentration F1.5v presented similar anticoagulant activity. The fucans F0.5v and F0.7v at 1.0 mg/mL showed high ferric chelating activity (~45%), whereas fucans F1.3v (0.5 mg/mL) showed considerable reducing power, about 53.2% of the activity of vitamin C. The fucan F1.5v presented the most prominent anticoagulant activity. The best antiproliferative activity was found with fucans F1.3v and F0.7v. However, F1.3v activity was much higher than F0.7v inhibiting almost 100% of HeLa cell proliferation. These fucans have been selected for further studies on structural characterization as well as in vivo experiments, which are already in progress / No presente estudo, seis popula??es de polissacar?deos sulfatados foram obtidas a partir da alga Dictyopteris delicatula (Lamouroux, 1809) e suas atividades anticoagulante, antioxidante e antitumoral avaliadas. Deste modo, as seis fra??es (F0.5v, F0.7v, F1.0v, F1.3v, F1.5v e F2.0v) foram obtidas por prote?lise seguida por fracionamento de acetona e gel filtra??o molecular Sephadex G-100. Todas as fra??es apresentaram atividade anticoagulante frente ao ensaio de aPTT, mas n?o sobre o ensaio de PT. As heterofucanas exibiram atividade antioxidante total, capacidade em sequestrar radicais super?xido e propriedade de quelar ferro. As an?lises qu?micas demonstraram que todos os polissacar?deos cont?m heterofucanas composta principalmente por fucose, xilose, glicose, galactose, ?cido ur?nico, e sulfato. Nenhuma das fra??es alterou o PT. Entretanto, todas as fra??es foram capazes de dobrar o aPTT de uma maneira dose dependente. As heterofucanas F0.7v e F1.0v demonstraram baixa atividade anticoagulante, enquanto a F1.5v apresentou a maior atividade anticoagulante e quando comparada com Clexane?, uma heparina de baixo peso molecular, em mesma concentra??o, apresentou antividade anticoagulante semelhante. As fucanas F0.5v e F0.7v a 1,0 mg/mL mostraram alta atividade de quela??o ferica (~ 45%), enquanto a fucana F1.3v (0,5 mg/mL) mostrou consider?vel potencial redutor, cerca de 53,2% da atividade da vitamina C. A melhor atividade antitumoral foi encontrada nas fucanas F1.3v e F0.7v. No entanto, a atividade F1.3v foi muito superior a F0,7v, quase 100% de inibi??o da prolifera??o de c?lulas HeLa (c?lula de c?ncer de colo de ?tero). Estas fucanas foram selecionadas para novos estudos sobre caracteriza??o estrutural, bem como em experimentos in vivo, que j? est?o em andamento / 2020-01-01
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Impacto do farelo do baga?o do caju na alimenta??o de su?nos dos 60 aos 90 Kg: estudo da digestibilidade dos nutrientes, desempenho e viabilidade econ?mica / The cashew pulp bran impact in swine finishing feeding (60 to 90 Kg): nutrition digestibility, performance and economic viability researchMota, Lorena Cunha 21 February 2014 (has links)
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Previous issue date: 2014-02-21 / Objetivou-se com este estudo avaliar a digestibilidade do farelo do baga?o de caju, o
desempenho de su?nos em termina??o alimentados com ra??es contendo n?veis crescentes
de farelo do baga?o do caju e a viabilidade econ?mica das ra??es. Foram realizados dois
experimentos. No primeiro experimento foi desenvolvido o ensaio de digestibilidade com
dez su?nos machos castrados pesando em m?dia 60?6,86kg, alojados em gaiolas
metab?licas por um per?odo experimental de quinze dias. Os animais foram alimentados
diariamente ? 8h00 e 15h00, sendo que cinco animais receberam a ra??o padr?o e os outros
cinco receberam a ra??o teste, com 30% de substitui??o da ra??o padr?o pelo FBC.
Realizou-se a coleta total de fezes sem uso de marcador, diariamente ?s 7h00, sendo
retirada uma al?quota de 20% e armazenada em freezer. Ao final do per?odo experimental
as amostras de fezes foram analisadas em laborat?rio para avaliar a digestibilidade da
mat?ria seca, prote?na bruta, fibra em detergente neutro, fibra em detergente ?cido, e os
teores de energia bruta, energia digest?vel, c?lcio e f?sforo do farelo do baga?o do caju. No
segundo experimento realizou-se o ensaio de desempenho e a viabilidade econ?mica das
ra??es, que foi composto por quarenta animais h?bridos comerciais (vinte machos castrados
e vinte f?meas), pesando em m?dia 60?5,24kg, delineados em blocos casualizados, com
cinco tratamentos contendo n?veis crescentes de farelo do baga?o do caju (0; 7,5; 15; 22,5
e 30 %) e quatro repeti??es. Os par?metros de desempenho avaliados foram o consumo
di?rio de ra??o, ganho di?rio de peso e convers?o alimentar. Os animais foram abatidos ao
atingiram peso de 90?6,8 kg, e dados da carca?a (espessura de toucinho, profundidade de
lombo, porcentagem da carne magra e peso da carca?a quente) foram coletados para
compor o estudo da viabilidade econ?mica, no qual, foram calculadas a receita bruta
parcial com ?ndice de bonifica??o, receita bruta parcial, receita l?quida parcial com ?ndice
de bonifica??o e receita l?quida parcial das ra??es. Os dados foram submetidos ? an?lise de
vari?ncia e regress?o por meio do programa estat?stico SAS. Observou-se que os
coeficientes de digestibilidade da mat?ria seca, prote?na bruta, fibra em detergente neutro e
fibra em detergente ?cido e os valores da energia bruta, energia digest?vel, mat?ria mineral,
Ca e P do FBC foram, 44,265%; 8,20%; 15,22%; 13,05%; 4.081 kcal/kg, 2.470 kcal/kg,
6,61%; 0,50%; 0,35%, respectivamente. A inclus?o de 7,5 % de farelo do baga?o de caju
incrementou a espessura de toucinho, porcentagem de carne magra e a receita bruta parcial
com ?ndice de bonifica??o / The research objective was to evaluate the cashew pulp bran (CPB)
digestibility, the finishing swine performance feed with different levels of cashew pulp
bran diets and the feed economic viability. Two experiments were conducted. In the first
experiment, the digestibility essay with ten barrows with initial average weight of 60?6.86,
housed in metabolic cages for a trial period of fifteen days was developed. The animals
were feed daily at 8h00 and 15h00. Five of this pigs received standard food and the other
five received test food, with 30 % of CPB replacement of the standard food. Was chosen
the total feces collection, without marker feces. The feces samples were collected daily at
7h00 and was taken at a rate of 20 % and stored in a freezer. At the end of the experimental
period, the feces samples were analyzed in the laboratory to evaluate the digestibility of
dry matter, crude protein, neutral detergent fiber, acid detergent fiber and the levels of
gross energy, digestible energy, Ca and P of the CPB. In the second experiment, was
conducted the performance essay and diets animals economic viability, with 40 crossbred
animals, weighing 60kg ?5,24 (20 castrated males and 20 females), outlined in randomized
block design with five treatments (0, 7.5, 15, 22.5, 30 % with CPB included) and four
replications. The parameters were the feed intake, weight gain, feed conversion. When
animals dyed average weight 90?6.8 kg were slaughtered. The carcass data (the backfat
depth, the longissimus muscle, the hot carcass weight and the lean meat percent) were
analyzed to compose the economic viability study, which evaluated the partial gross
receipts with rate bonus, partial gross receipts, partial liquid receipts and partial liquid
receipts with rate bonus. The parameters were subjected to variance and regression
analysis using the SAS program. It was observed, the dry matter, crude protein, neutral
detergent fiber and acid detergent fiber digestibility and gross energy, digestible energy,
ash, Ca and P cashew pulp bran values were, 44.265% ; 8.20%; 15.22%; 13.05%; 4,081
kcal / kg 2,470 kcal / kg, 6.61%; 0.50%; 0.35%, respectively. The diet with 7.5% increased
the backfat depth, the lean meat percentage and the partial gross revenue allowance index
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Caracteriza??o parcial e atividades farmacol?gicas do extrato rico em polissacar?deos sulfatatos da angiosperma marinha Halodule wrightiiCosta, Leandro Silva 25 November 2008 (has links)
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Previous issue date: 2008-11-25 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Sulfated polysaccharides (PS) are biomolecules with a great biotechnological potential. There are few data about PS from high plants. In addition, pharmacological activities of PS from plants have not been carrying out. The aim of this work was extract PS from the angiosperm Halodule wrightii and study their anticoagulant and antioxidant activities. Histological analysis showed the presence of the PS manly in the roots. A polysaccharide-rich extract was obtained from H. wrightii by proteolysis followed by methanol and TCA precipitation. Chemical, infra-red analysis and agarose gel electrophoresis in 1.3 diaminopropane acetate buffer confirmed the presence of sulfated polysaccharides made by glucose, galactose, xylose and sulfate residues in the proportion 1: 0,9: 1: 1. In addition polyacrilamide electrophoresis have shown that extract is mainly compose by 11kDa sulfated polysaccharides. Pharmacological analysis have shown total antioxidant capacity (CAT) that resulted in 15,21 μg for equivalent of ascorbic acid, scavenging activity of the DPPH radical with 41,36 % of scavenging, activity of reducing power with the maximum of 0,290 nm (50 % of vitamin C activity) and scavenging activity superoxide radical (O2-) with a maximum of 32,23 %. Chelating activity of metal less than 4% and scavenging activity of the radical hydroxyl (OH-) less than 2%. Time of activated partial tromboplastin (aPTT) doubling the time of coagulation from 20μg of and protrombin time (PT) was not present. The data indicate that PS from Halodule wrightii could be considered for future applications in medicine, food production or cosmetic industry / Os polissacar?deos sulfatados (PS) s?o biomol?culas com um grande potencial biotecnol?gico por apresentarem uma diversidade estrutural e farmacol?gica muito grande. Poucos s?o os relatos da exist?ncia destes pol?meros em vegetais superiores, al?m disso, ainda n?o se tem relatos da identifica??o de atividades antioxidantes e anticoagulantes com PS extra?dos destes vegetais. Com o intuito de verificar a presen?a destes polissacar?deos em angiospermas marinhas conhecidas popularmente como capim do mar, foi coletada a esp?cie marinha Halodule wrightii. As por??es vegetativas (folha, caule e raiz) n?o foram separadas sendo preparado inicialmente um extrato denominado de extrato bruto (EB). Ap?s descontamina??o prot?ica o material obtido foi chamado de extrato de polissacar?deos totais (EPT). A presen?a destes polissacar?deos foi investigada e confirmada por an?lises qu?micas, espectroscopia de infravermelho e eletroforese em gel de agarose sendo denominados de extrato rico em polissacar?deos sulfatados (EPS). A an?lise histol?gica da localiza??o dos PS resultou na presen?a destes polissacar?deos principalmente na epiderme da por??o vegetativa raiz. As an?lises qu?micas mostraram que os polissacar?deos contem glicose, galactose, xilose e sulfato na propor??o de 1: 0,9: 1: 1 e massa molecular de aproximadamente 11 kDa. Os grupos sulfatos est?o provavelmente ligados principalmente em C2 de um monossacar?deo. Testes de atividade antioxidante demonstraram que os PS de H. wrightii n?o apresentaram resultado para o teste de capacidade antioxidante total pelo m?todo CAT. Desta forma foi utilizada a atividade de seq?estro do radical DPPH indicado na literatura por dosar a capacidade antioxidante total que resultou em 41,36%. O seq?estro do ?on super?xido tamb?m foi realizado e resultou em 32,23% assim como o poder redutor que equivaleu a 50% da atividade da Vit. C. N?o houve atividade de seq?estro do radical hidroxila assim como atividade quelante de metal. O teste de atividade anticoagulante (aPTT) mostrou que EPS dobra o tempo de coagula??o com 20 g, que ? apenas 2,5 vezes a quantidade da Clexane? (heparina de baixo peso molecular). Para o tempo de protrombina (PT) H.wrightii n?o apresentou atividade. Os dados indicam que EPS possuem um potencial biotecnol?gico anticoagulante e antioxidante e que futuras an?lises se fazem necess?rias para confirmarem esse potencial
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