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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
131

Efeitos do azul de metileno na lesão pulmonar aguda induzida por ácido oleico em ratos / Effects of methylene blue in acute lung injury induced by oleic acid in rats

Ana Paula Cassiano Silveira 10 June 2014 (has links)
INTRODUÇÃO. O termo Lesão Pulmonar Aguda (LPA) é usado para descrever a resposta pulmonar à lesão que ocorre diretamente ou indiretamente nos pulmões. A quebra da barreira alvéolo-capilar determina o influxo de líquido rico em proteínas para dentro dos espaços alveolares, sendo necessária a reabsorção desse líquido no processo de resolução da LPA. A infusão intravenosa de Ácido Oleico (AO) em ratos provoca agudamente edema alveolar difuso e focos hemorrágicos intra-alveolares, sendo um bom modelo de indução. Estudos relatam que o Azul de Metileno (AM) atenua tais lesões, com efeito protetor, no tecido pulmonar, e reduz o edema presente na LPA em animais com sepse através da inibição da guanilato ciclase solúvel (GCs), uma enzima ativadora da via NO-GMPc. OBJETIVO. Estudar a repercussão da inibição da GCs pelo AM na permeabilidade capilar pulmonar ministrando-o antes e após a indução da lesão pulmonar por AO. MÉTODO. Ratos Wistar foram divididos em 5 grupos: Sham com infusão de salina em bolus; AM com infusão de AM por 2h; AO com infusão de AO em bolus, AM/AO com infusão de AM por 2h, sendo que, após 5 min do início, recebeu AO simultaneamente em bolus e AO/AM com infusão de AO em bolus e, após 2h, infusão de AM por mais 2h. Após 4h foi realizada a coleta de materiais (sangue, lavado bronco-alveolar e tecido pulmonar) para análise do NO plasmático e tecidual, gasometria arterial, cálculo do peso úmido/peso seco (PU/PS) e histologia do tecido pulmonar de todos os grupos. A estatística utilizada foi a análise de variância (one-way ANOVA) com p<0.05. RESULTADO. Não foi encontrado hipoxemia grave após 4h de lesão. O grupo AO apresenta um aumento no número de proteínas no lavado bronco-alveolar e na relação PU/PS comparado aos grupos controle: Sham e AM, confirmando a presença de lesão e alteração de permeabilidade pulmonar. Os grupos tratados com AM apresentaram melhora na permeabilidade pulmonar, porém, apenas o grupo pré-tratamento (AM/AO) apresentou diferença estatística na redução do extravasamento de proteínas no lavado. Não foram encontradas diferenças significativas no NO plasmático e tecidual. Na microscopia, a congestão capilar foi intensa, acompanhada de múltiplos focos de edema alveolar, exsudato intra-alveolar proteico, áreas de hemorragia e infiltrado inflamatório neutrofílico, tanto no interstício quanto nos septos alveolares.Os grupos tratados com AM apresentaram diminuição das áreas de edema, exsudação e hemorragia, porém, com maior evidência no grupo AM/AO. CONCLUSÃO. O AM diminui a permeabilidade pulmonar quando administrado de maneira precoce amenizando os danos causados pela LPA induzida por AO. / BACKGROUND. The term Acute Lung Injury (ALI) is used to describe the response to lung injury that occurs directly or indirectly in the lungs. The rupture of the alveolar - capillary barrier determines the influx of protein-rich fluid into the alveolar spaces, the reabsorption of this fluid in the process of resolving the ALI is required. Intravenous infusion of oleic acid (OA) in rats acutely causes diffuse alveolar edema and intra-alveolar hemorrhagic foci, being a good role model induction. Studies have reported that Methylene Blue (MB) attenuates such injuries, with a protective effect in lung tissue and reduce edema present in the ALI present in with sepsis by inhibition of soluble guanylate cyclase (sGC), an activator of the enzyme NO- cGMP pathway. OBJECTIVE. To study the effect of inhibition of sGC by MB in pulmonary capillary permeability ministering to the before and after induction of lung injury by OA. METHOD. Wistar rats were divided into 5 groups: Sham infused with saline bolus, MB infused with MB for 2hrs, OA infused with OA bolus, MB/OA infused with MB for 2hrs, and after 5 min from the beginning, simultaneously received OA bolus and OA/MB infused with OA bolus and after 2hrs, MB infusion for 2hrs. After 4hrs the collection of materials was performed (blood, bronchoalveolar lavage and lung tissue) for analysis of plasma and tissue NO, arterial blood gases, calculation of the wet weight/dry weight (WW/DW) and histology of lung tissue from all groups. The statistic used was analysis of variance (one-way ANOVA) with p<0.05. RESULTS. Not found severe hypoxemia after 4hrs of injury. The OA group shows an increase in the number of proteins in bronchoalveolar lavage and in WW/DW ratio compared to the control groups: Sham and MB, confirming the presence of injury and alterations of lung permeability. The groups treated with MB showed improvement in lung permeability, however, only the pretreatment group (MB/OA) showed statistical significance in reducing the leakage of protein in the lavage. No significant differences were found in plasma and tissue NO. In microscopy, capillary congestion was intense, accompanied by multiple foci of alveolar edema, intra-alveolar proteinaceous exudates, areas of hemorrhage and neutrophilic inflammatory infiltrate in both the interstitium and in the alveolar septa. The groups treated with MB showed reduction in areas of edema, exudation and hemorrhage, however, most obviously in MB/OA group. CONCLUSION. The MB decreases lung permeability when administered as early as possible, mitigating the damage caused by OA-induced ALI.
132

Eficiência fotodinâmica dos fenotiazínicos em mitocôndrias e células tumorais / Photodynamic efficiency of phenotiazinic compounds into mitochondria and tumor cells

Dino Santesso Gabrielli 14 June 2007 (has links)
O mecanismo de ação de fotossensibilizadores (FS) fenotiazínicos (azul de metileno - AM, tionina - TIO e azure B - AB) foi estudado em mitocôndrias e células Hela, e a dosimetria de AM em células HL60. Foi observado que a ligação de AM em mitocôndrias é dependente da concentração de mitocôndrias, concentração de AM e do potencial de membrana mitocondrial (&#916; &#936;). A geração de oxigênio singlete a partir de AM é influenciada pelo &#916; &#936;, sendo aproximadamente o dobro em mitocôndrias desacopladas. Essa variação se deve a influência do &#916; &#936; sobre a relação dímero/monômero (D/M) do AM. A ligação de AM também é reduzida pela metade em mitocôndrias desacopladas. Esses efeitos não foram observados para os outros fotossensibilizadores, embora tenham a mesma carga e estrutura semelhante. Isso indica um efeito diferencial do &#916; &#936; sobre o AM. Foi observada a toxicidade desses FSs em células Hela. Sem irradiação, apenas AB se mostrou tóxico. Em amostras irradiadas, AB e AM se mostraram tóxicos. Todos os FSs mostraram capacidade de produzir núcleos sub-diplóides, típicos de apoptose, com a maior eficiência sendo do AB. Em relação a toxicidade no escuro e claro, AB mostrou toxicidade em ambas as condições, TIO mostrou pouca toxicidade em ambas as condições e AM mostrou pouca toxicidade no escuro e alta toxicidade quando irradiado. A dosimetria que favorece apoptose em células HL60 também foi testada, sendo que µM de AM, os melhores resultados para formação de núcleos sub-diplóides foram com 10 2 com irradiação (contínua ou fracionada) de 0,55 J/cm2 . Essas dosagens foram as mais baixas utilizadas, indicando que um aumento na concentração de FS ou de luz tende a levar as células à morte necrótica. Conclui-se que os FSs fenotiazínicos tem afinidade por mitocôndrias e por células, podendo levar a indução de apoptose em doses não tóxicas no escuro. / The mechanism of action of phenotiazinic fotossensitizers (FS) (methylene blue - AM, tionin - TIO and azure B - AB) was studied into mitochondria and Hela cells, and AM dosimetry into HL60 cells was determined. AM binding to mitochondria is dependent of mitochondrial concentration, AM concentration and mitochondrial membrane potentials (&#916; &#936;). Singlet oxygen generation from AM is influenced by &#916; &#936;, almost doubling in uncoupled mitochondria. This effect is due to the influence of &#916; &#936; on AM dimer/monomer (D/M) ratio. Also, AM binding drops to half into uncoupled mitochondria. For all the others FSs studied, these effects were not observed, even though they present the same &#916; &#936; action over AM. charge and similar structure. This indicates a specific FSs toxicity was observed in Hela cells. In the dark, only AB showed toxicity. In irradiated samples, AM and AB showed toxicity. All the tested FSs showed capacity to induce sub-diploid nuclei formation, a hallmark of apoptosis, with AB showing the highest efficiency. AB showed high toxicity in both (irradiated and non-irradiated) conditions, TIO showed low toxicity in both conditions and AM showed low toxicity in the dark and high toxicity with laser irradiation. The dose that promotes apoptotic cell death in HL60 cells was also tested. Results 2 were observed at 10 µM AM, with laser irradiation (fraction or continuous) of 0,55 J/cm . These doses were the lowest doses tested, indicating that high doses of FS or laser induce necrotic cell death. We conclude that phenotiazinic FSs have affinity for mitochondria and cells, and induce apoptosis at doses that are not toxic without irradiation
133

Desenvolvimento de nanopartículas fotossensibilizadoras / Development of photosensitizing nanoparticles

Dayane Batista Tada 14 December 2007 (has links)
No presente trabalho são apresentadas a síntese e a caracterização estrutural, fotofísica, fotoquímica e fotobiológica de nanopartículas contendo os fotossensibilizadores (FS) Azul de Metileno (AM) e Tionina. AM e Tionina foram incorporados nas nanopartículas sil-AM e sil-Tio pelo processo sol-gel. Nas nanopartículas Cab-Tio, Tionina foi ligada à superfície de sílica CabOsil® através de ligação covalente com reagentes bifuncionais. Todas as nanopartículas mostraram-se esféricas e com de diâmetro médio na faixa de 30 a 60nm. A imobilização dos FS induziu a agregação destes em extensões diferentes para cada tipo de nanopartícula. Foi observado que a maior presença de dímeros de FS leva à menor eficiência de geração de 1O2. Constatou-se que as nanopartículas sofrem pouca influência do meio, uma vez que os FS a elas ligadas não sofreram redução química por NADPH, nem supressão do estado tripleto por íons ascorbato e a supressão de fluorescência por íon brometo foi diminuída. Foi testado também o efeito do recobrimento destas nanopartículas com lipídios dioleilfosfatidil colina (DOPC) e fosfatidilglicerol (PG) e com Polietileno glicol (PEG). A adsorção das nanopartículas sobre membranas miméticas foi reduzida após os recobrimentos, resultado que foi explicado pelas interações de carga superficial (potencial zeta) e pela força de hidratação. As nanopartículas sil-AM e Cab-Tio apresentaram fototoxicidades in vitro, 38% e 20% maiores que os respectivos FS livres. A modificação das nanopartículas de sil-AM com lipídios e com PEG diminuiu a fototoxicidade das mesmas e no caso do recobrimento com lipídios levou ao aumento da toxicidade no escuro. Imagens de microscopia confocal mostraram que as nanopartículas com e sem recobrimento de lipídios entram em células B16. No caso das nanopartículas recobertas, observou-se um perfil de distribuição difuso por todo o citoplasma e no caso de nanopartículas sem recobrimento, estas encontraram-se em poucas regiões vacuolares do citoplasma. O perfil de distribuição homogênea por todo o citoplasma no caso de nanopartículas recobertas com lipídios pode ser o responsável pelo aumento de toxicidade no escuro. Concluiu-se que a ligação dos FS em nanopartículas com diferentes graus de agregação pode ser uma estratégia para obtenção de sistemas com capacidade modulada de geração de 1O2 e com reduzida susceptibilidade às composições do meio. As atividades fototóxicas das nanopartículas contra células B16 mostraram que estas podem ser úteis em Terapia Fotodinâmica de Câncer / In this work we present the synthesis and the characterization (structural, photophysical, photochemical and photobiological) of nanoparticles with incorporated photosensitizers (PS) Methylene Blue (MB) and Thionin. MB and Thionin were incorporated in sil-MB and sil-Th nanoparticles through sol-gel process. In the case of Cab-Th nanoparticles Thionin was linked to the surface of CabOsil® nanoparticles through cross-linking reactions. All nanoparticles were spherical and presented average diameter in the range of 30 to 60nm. Different extension of PS aggregation was observed in each nanoparticle. It was characterized that the higher the proportion of dimers to monomers the smaller the efficiency of singlet oxygen (1O2) generation. It was shown that nanoparticles can protect PS from external interferences, since NADPH did not reduce them, neither were their triplet state quenched by ascorbate ions. Besides, fluorescence quenching by bromide ions was reduced compared to free PS. The effect of covering the nanoparticles with lipids, i.e., di-oleil phosphatidylcholine (DOPC) and phosphatidylglycerol (PG), and with Polyethylene glycol was also tested. The nanoparticle adsorption over membrane mimics was reduced, which was explained by the interaction among surface charges (zeta potential) and by hydration forces. Sil-MB and Cab-Th nanoparticles presented in vitro phototoxicity 38% and 20% higher than the respective free PS. It was observed that the nanoparticle coating with lipids and with PEG reduced their photoxicity. Nanoparticles coated with lipids showed higher toxicity in the dark. Confocal fluorescence images of B16 cells showed that nanoparticles with or without lipid coating enter the cells. In the case of lipid-coated nanoparticles a diffuse distribution profile was observed and in the case of nanoparticles without coating, they concentrated in specific vacuolar regions of the cytoplasm. The homogeneous cytoplasmic distribution profile of lipid-coated nanoparticles can explain the increased toxicity in the dark. It has been concluded that immobilization of PS with different aggregation degrees is a strategy to obtain systems in which the modulated efficiency of 1O2 generation is not affected by the external medium. Finally, based on the observed in vitro phototoxicity activity against B16 cells, these systems can be useful in Photodynamic Therapy of Cancer
134

Hydrogely s uhlíkovými vlákny / Hydrogels with carbon fibers

Kučerová, Barbora January 2021 (has links)
This diploma thesis is focused on the study of the behaviour of carbon fibers in the hydrogel, especially the viscoelastic modules od these gels ort he coefficient of color transmission through the gel. The main goals were to optimize the samples and measure their viscoelastic modules on a rheometer, in several different modes. This was followed by simulation of physiological conditions for selected samples and their measurement on the rheometer. There was also an experiment of continuous diffusion in cuvettes, in which the penetration of the dye through the gel with the addition of fibers and for comparison also without them. Last but not least diffusion pairs were prepared.
135

Stanovení tenzidů v odpadech a odpadních vodách / Determination of tensides in waste and waste waters

Sigetová, Martina January 2013 (has links)
Surfactants belong to the surface active agents with the ability to reduce surface tension, which is used mainly in detergents and cleaners. The diploma thesis is focused on the determination of surfactants in waste and waste waters. The theoretical part describes the basic properties of surfactants, their distribution, valid legislation in the EU and SR in the field of working with these materials and a variety of analytical methods for the determination of these substances. The conclusion of the thesis is to evaluate the results of measuring the concentration of anionic surfactants, which were determined in samples of waste water and sludge.
136

Composite Nanostructures as Effective Catalysts for Visible-Light-Driven Chemical Transformations

Rasamani, Kowsalya Devi, 0000-0002-1717-1426 January 2020 (has links)
The development of nanoscale heterostructure photocatalysts for the effective, direct utilization of visible light (400-750 nm, ~44% of solar spectrum) to drive important chemical conversions is a prime research area in the field of photocatalysis. Particles at nanoscale dimensions have a large surface area-to-volume ratio, expose a high number of active surface sites, and exhibit unique electronic properties (different from bulk) that are beneficial for improving the overall catalytic activity. However, the advantages of size reduction are often overshadowed by the low optical absorption (as absorption power  size3) and colloidal instability (extensive aggregation) of particles at the nanoscale. In this dissertation, we demonstrate a strategy to improve the colloidal stability and enhance the optical absorption of nano-sized semiconductor and metal nanoparticles (NPs) that exhibit weak visible light absorption. The colloidal, free-standing NPs are placed on transparent, dielectric silica nanospheres (SiOx NSs) that act as optical antenna supports, forming SiOx/NP composite nanostructures. The spherical morphology of SiOx enables scattering resonances (Fabry Perot or Whispering Gallery Modes) which enhances the local electric field on or near the surface of the NS. The NPs placed on the surface of SiOx NS interact with the locally enhanced electric field and exhibit improved optical absorption. By varying the size of the SiOx NS, the resonance wavelengths and the intensity of the local electric field enhancement can be tuned, offering the ability of such structures to effectively utilize a wide range of energies in the visible region. Composite nanostructures comprised of various classes of nanomaterials such as metal-doped semiconductor, plasmonic, and non-plasmonic metal NPs were investigated to perform the desirable solar-to-chemical transformations. First, we employed SiOx-loaded silver-doped silver chloride (SiOx/AgCl(Ag)) photocatalyst to investigate the role of metal-induced gap states in AgCl, a wide bandgap semiconductor. SiOx/AgCl(Ag) exhibit high catalytic performance and photostability after 10 cycles of the probe reaction, methylene blue (MB) degradation under visible light irradiation. The results indicate that the visible light absorption due to metal-induced gap states can be further improved by employing the SiOx NSs as supports that act as optical nanoantenna. We then studied the influence of NP size on the catalytic activity to understand the effect of size in promoting the generation and transfer of hot electrons to surface adsorbates. Our findings indicate that upon employing Ag NPs of different particle size (<10 nm and >10 nm) and normalizing for the optical absorption and moles of surface Ag atoms, the efficient generation and transfer of photoexcited hot electrons is favored in the small-sized Ag NPs (size <10 nm) than bigger Ag NPs. Next, we investigated the selective partial hydrogenation of nitroarene to N-aryl hydroxylamine using SiOx-loaded platinum (SiOx/Pt) photocatalysts. We found that change in the surface electronic structure of the small Pt NPs (size <5 nm) due to light illumination and surface modification (by adding suitable organic ligands), minimize the adsorption of the electron-rich hydroxylamine molecules and minimize their complete conversion to aniline, resulting in high N-hydroxylamine selectivity. Overall, our work shows that well-controlled composite nanostructures comprising of active catalyst loaded on dielectric SiOx NS supports that act as optical nanoantenna are a promising class of photocatalysts for driving photon-to-chemical transformations with high activity and product selectivity. / Chemistry
137

Rhamnolipid Biosurfactant Production from Glycerol: New Methods of Analysis and Improved Denitrifying Fermentation

Pinzon-Gamez, Neissa M. 15 December 2009 (has links)
No description available.
138

Biogenic Nanosilica Synthesis Employing Agro-Waste Rice Straw and Its Application Study in Photocatalytic Degradation of Cationic Dye

Singh, Garima, Dizaji, Hossein Beidaghy, Puttuswamy, Hariprasad, Sharma, Satyawati 02 June 2023 (has links)
The current study aims towards a holistic utilization of agro-waste rice straw (RS) to synthesize nanosilica (SiNPs) employing the sol–gel method. The effect of ashing temperature was evaluated for the synthesis process. X-ray diffraction demonstrated a broad spectrum at 21.22° for SiNPs obtained using RSA-600, signifying its amorphous nature, whereas crystalline SiNPs were synthesized using RSA-900. The EDX and FTIR spectra confirmed the significant peaks of Si and O for amorphous SiNPs, confirming their purity over crystalline SiNPs. FE-SEM and TEM micrographs indicated the spheroid morphology of the SiNPs with an average size of 27.47 nm (amorphous SiNPs) and 52.79 nm (crystalline SiNPs). Amorphous SiNPs possessed a high surface area of 226.11 m2/g over crystalline SiNPs (84.45 m2/g). The results obtained attest that the amorphous SiNPs possessed better attributes than crystalline SiNPs, omitting the need to incorporate high temperature. Photocatalytic degradation of methylene blue using SiNPs reflected that 66.26% of the dye was degraded in the first 10 min. The degradation study showed first-order kinetics with a half-life of 6.79 min. The cost-effective and environmentally friendly process offers a sustainable route to meet the increasing demand for SiNPs in industrial sectors. The study proposes a sustainable solution to stubble burning, intending towards zero waste generation, bioeconomy, and achieving the Sustainable Development Goals (SDGs), namely SDG 13(Climate Action), SDG 3(Good health and well-being), SDG 7(use of crop residues in industrial sectors) and SDG 8 (employment generation).
139

Évaluation de l’ototoxicité secondaire au bleu de méthylène chez le cochon d’Inde : étude animale prospective

Belhassen, Sarah 04 1900 (has links)
INTRODUCTION: Le bleu de méthylène est un colorant grandement utilisé dans le domaine médical, notamment pour ses propriétés de coloration histologique. Il est également utilisé comme agent photosensibilisant dans la thérapie photodynamique antimicrobienne, qui une fois photoactivé devient efficace pour l’éradication de plusieurs germes multirésistants. L’objectif de cette étude est d’investiguer le potentiel ototoxique du bleu de méthylène. MÉTHODES: Vingt cochons d’Inde divisés en deux groupes, ont reçu une solution de bleu de méthylène et de gentamicine dans l’oreille testée. L’oreille controlatérale a reçu une solution saline contrôle. Nous avons procédés à des potentiels évoqués auditifs du tronc cérébral avant et une semaine suivant la série d’injections. À la suite des dissections, des analyses histologiques et immunohistochimiques ont été réalisées. RÉSULTATS: La différence moyenne de perte auditive dans le groupe gentamicine comparativement au groupe normal salin était de 66.25 dB (p<0.001). Toutefois, la différence moyenne de perte auditive dans le groupe ayant reçu du bleu de méthylène comparativement à celui ayant reçu des injections de la solution saline était de 1.50 dB, et n’a pas été démontré comme étant statistiquement significative (p=0.688). De plus, la captation de caspase-3 en immunohistochimie (marqueur d’apoptose) n’a pas été significative dans le groupe recevant le bleu de méthylène. CONCLUSION: À la lumière de nos résultats, les injections intratympaniques de bleu de méthylène n’ont pas démontrées de potentiel ototoxique. Nous recommandons des études supplémentaires afin d’en préciser son utilisation sécuritaire dans le domaine otologique. / INTRODUCTION: Methylene blue is widely used in the medical field, especially as a blue dye for staining. It is also used as a photosensitizing agent in antimicrobial photodynamic therapy, which once photoactivated is effective for the eradication of several multiresistant bacteria. The objective of this study is to investigate its ototoxic potential. METHODS: Twenty guinea pigs, forming two groups, received respectively intratympanic methylene blue and gentamicin in one ear. The contralateral ears received a control saline solution. We conducted auditory evoked brainstem response (ABR) before and one week after the injection series. Once completed, the cochleas were dissected and analyzed by histology and immunohistochemistry. RESULTS: The mean difference of hearing loss in the saline group compared to the gentamicin group was 66.25dB (p<0.001). However, the mean difference of hearing loss in the methylene blue group compared to normal saline was 1.50 dB, and it was not shown to be statistically significant (p=0.688). Furthermore, uptake of caspase-3 by immunohistochemistry (apoptotic marker) was negative in the group which received injections of methylene blue. CONCLUSION: In light of our results, intratympanic injections of methylene blue did not demonstrate an ototoxic potential. We recommend further studies to precise its usefulness in the otology field.
140

Estudo de propriedades biofísicas de membrana sob estresse oxidativo e a interação com proteínas formadoras de poros / Study of biophysical properties in membranes under oxidative stress and interaction with pore-forming proteins

Checchia, Robert Garcia 12 February 2019 (has links)
Neste trabalho investigamos efeitos de fotoirradiação e toxinas sob membranas celulares miméticas. Foram utilizadas, como modelo de membranas lipídicas, vesiculas unilamelares gigantes (GUVs) compostas pro lipídeos oxidados e não oxidados observadas por microscopia ótica de contraste de fase. Inicialmente estudamos a foto-resposta de membranas compostas por POPC e POPG dispersas em solução contendo azul de metileno (MB). Na sequência, estudamos o efeito de toxinas formadoras de poros, Esticolisina I (ST I) e Esticolisina II (ST II), em membranas contendo lipídeos oxidados e não oxidados. Os resultados de MB (10 µM) disperso em solução de membranas compostas por POPC e o lipídeo aniônico POPG indicaram que o aumento da densidade de carga negativa nas membranas das GUVs, que favorece a ligação da moléculas positivamente carregadas como MB nas membranas, tem como consequência um aumento de permeabilidade da membrana muito mais rápído em relação a membranas compostas apenas por POPC. Isto se deve ao fato que a localização preferencial do MB na membrana de POPC:POPG favorece a formação de oxigênio singlete próximo a dupla ligação da cadeia alquílica, dando início a reação de peroxidação lipídica de maneira mais efetiva que em membrana de POPC. Os resultados da ação das toxinas STI e STII (21 nM) em GUVs contendo lipídeos não oxidados PC e esfingomielina evidenciam que apenas STII é capaz de permear estas membranas a esta concentração. Mais ainda, nossos resultados sugerem que a existência de separação de fases fluida-gel na bicamada lipidica composta por PC:SM (razão molar 1:1) favorece a ação da toxina StII. Ao analisarmos membranas contendo lipídeos hidroperoxidados (POPC-OOH) dispersas em solução contendo STII (21 nM) observamos um aumento de permeabilidade na membrana num conjunto de GUVs, associado a formação de poros, apenas em bicamadas lipídicas formadas por misturas de lipídeos oxidados (POPC) e não oxidados (POPC-OOH). Quanto maior a concentração de lipídeos oxidados na membrana mais rapidamente ocorre o aumento de permeabilidade. / In this work we investigate the effects of photoirradiation and toxins on mimetic cell membranes. As a model of lipid membranes, giant unilamellar vesicles (GUVs) composed of oxidized and oxidized pro-lipids were observed by optical phase contrast microscopy. Initially we studied the photo-response of membranes composed of POPC and POPG dispersed in solution containing methylene blue (MB). Following, we studied the effect of pore-forming toxins, Sticolysin I (ST I) and Sticolysin II (ST II), on membranes containing oxidized and non-oxidized lipids. The results of MB (10 M) dispersed in solution of membranes composed of POPC and the anionic lipid POPG indicated that the increase in the negative charge density in the membranes of GUVs, which favors the binding of positively charged molecules as MB in the membranes, consequently increases membrane permeability in regard to membranes composed only of POPC. This is due to the fact that the preferred location of the MB in the POPC: POPG membrane favors the formation of singlet oxygen near the double bond of the alkyl chain, initiating the lipid peroxidation reaction more effectively than in the POPC membrane. The results of the action of the STI and STII toxins (21 nM) on GUVs containing non oxidised lipids PC and sphingomyelin show that only STII is able to permeate these membranes at this concentration. Moreover, our results suggest that the existence of fluid-gel phase separation in the lipid bilayer composed of PC:SM (molar ratio 1:1) favors the action of the StII toxin. When analyzing membranes containing hydroperoxidized lipids (POPC-OOH) dispersed in solution containing STII (21 nM) we observed an increase in membrane permeability in a set of GUVs, associated with pore formation, only in lipid bilayers formed by mixtures of oxidized lipids (POPC-OOH) and non-oxidized ones. The higher the concentration of oxidized lipids in the membrane, the faster the permeability increases.

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