Avaliação da atividade antibacteriana e moduladora de aminoglicosídeos do óleo essencial de Cymbopogon citratus (DC.) Stapf

Vários trabalhos vêm demonstrando a importância do estudo de produtos naturais como fonte alternativa para novos antimicrobianos ou que venham potencializar os já existentes. Neste contexto este trabalho teve como objetivo investigar a atividade antibacteriana e as possíveis interações entre o óleo essencial de Cymbopogon citratus combinados a aminoglicosídeos frente a linhagens padrões e multirresistentes de S. aureus, E. coli e de P. aeruginosa provenientes de isolados clínicos. Um ensaio de microdiluição foi realizado para verificar a atividade antibacteriana e as possíveis interacções entre o produto natural e os antibióticos, utilizando uma concentração sub-inibitória. Através dos resultados foi constatado a interferência sinérgica dos aminoglicosídeos quando associados com o óleo essencial em uma concentração de CIM/8, com redução das CIMs em até quatro pontos frente às linhagens de S. aureus 358, E. coli 27 e P. eruginosa- 143. Mas nenhuma atividade modificadora foi observada frente a P. aeruginosa 78 e P. aeruginosa 91. Através dos resultados pode-se concluir que o óleo essencial de Cymbopogon citratus pode ser uma fonte alternativa de produtos naturais com atividade antibacterianaVarios trabajos están demostrando la importancia del estudio de productos naturales como una fuente alternativa para nuevos antimicrobianos o que puedan mejorar los ya existentes. En este contexto, el objetivo de este trabajo fue investigar la actividad antibacteriana y las posibles interacciones entre el aceite esencial de Cymbopogon citratus aminoglucósido combinado frente de linajes y patrones de S. aureus multirresistente, E. coli y P. aeruginosa provenientes de aislamientos clínicos. Se realizó una prueba de microdilución para verificar la actividad antibacteriana y posibles interacciones entre el producto natural y antibióticos, usando una concentración sub-inibitória. A través de los resultados se observan interferencia sinérgica de los aminoglucósidos cuando se asocian con el aceite esencial en una concentración de CIM/8, con reducción de CIMs de hasta cuatro puntos contra las cepas de S. aureus 358, E. coli 27 y P. aeruginosa 143. Pero no se observó ninguna actividad modificadora contra P. aeruginosa 78 y P. aeruginosa 91. A través de los resultados se puede concluir que el aceite esencial de Cymbopogon citratus puede ser una fuente alternativa de productos naturales con actividad antibacterianaSeveral works demonstrated the importance of the study of natural products as an alternative source for new antimicrobial drugs or for modulators for these ones. In this point, the aim of this was to investigate the antibacterial activity and the possible interactions between the essential oil of Cymbopogon citratus alone and in association with aminoglycosides against standard and clinically isolated strains of multidrug-resistant bacteria such as S. aureus, E. coli and P. aeruginosa by microdilution method. The results indicated a synergism between the antibiotics and the essential oil with a subinhibitory concentration (MIC/8), reducing the minimal inhibitory concentration (MIC) sixteen times against the multidrug-resistant strains of S. aureus 358, E. coli 27 and P. aeruginosa 143, but none modulatory activity was observed against P. aeruginosa 78 and P. aeruginosa 91 strains. By our results, can be concluded that the essential oil of Cymbopogon citratus can be an interesting source of natural products with antibacterial and/or modulatory antibiotic activities

Essential Oil from <i>Croton blanchetianus</i> Leaves: Anticandidal Potential and Mechanisms of Action

Antimicrobial drugs are becoming ineffective given the resistance acquired by microorganisms. As such, it is imperative to seek new antimicrobial molecules that could provide a basis for the development of new drugs. Therefore, this work aimed to evaluate the antimicrobial potential and the mechanisms of action of the essential oil extracted from leaves of Croton blanchetianus (named CbEO) on different fungi and bacteria of clinical importance in both planktonic and biofilm lifestyles. GC-MS/MS analysis revealed the presence of twenty-two different compounds in the CbEO, which were identified using the Kovats retention index. Among these, the most abundant were amorphene (20.03%), spathulenol (5%), bicyclogermacrene (1.49%), caryophyllene oxide (4.55%), and eucalyptol (5.62%). CbOE (50 µg mL−1) barely inhibited the growth of Bacillus subtilis (23%), Pseudomonas aeruginosa (27%), and Salmonella enterica (28%), and no inhibition was obtained against Enterobacter aerogenes and Klebsiella pneumoniae. Additionally, no activity against bacterial biofilm was detected. In contrast, CbEO was active against Candida species. C. albicans and C. parapsilosis were inhibited by 78 and 75%, respectively. The antibiofilm potential also was favorable against C. albicans and C. parapsilosis, inhibiting 44 and 74% of biofilm formation and reducing around 41 and 27% of the preformed biofilm, respectively. CbOE caused membrane damage and pore formation, overproduction of ROS, and apoptosis on C. albicans and C. parapsilosis cells, as well as not inducing hemolysis in human red cells. The results obtained in this work raise the possibility of using the essential oil of C. blanchetianus leaves as an alternative to fight infections caused by C. albicans and C. parapsilosis

Vaccination With Recombinant Filamentous fd Phages Against Parasite Infection Requires TLR9 Expression

Recombinant filamentous fd bacteriophages (rfd) expressing antigenic peptides were shown to induce cell-mediated immune responses in the absence of added adjuvant, being a promising delivery system for vaccination. Here, we tested the capacity of rfd phages to protect against infection with the human protozoan Trypanosoma cruzi, the etiologic agent of Chagas Disease. For this, C57BL/6 (B6) and Tlr9−/− mice were vaccinated with rfd phages expressing the OVA257–264 peptide or the T. cruzi-immunodominant peptides PA8 and TSKB20 and challenged with either the T. cruzi Y-OVA or Y-strain, respectively. We found that vaccination with rfd phages induces anti-PA8 and anti-TSKB20 IgG production, expansion of Ag-specific IFN-γ, TNF-α, and Granzyme B-producing CD8+ T cells, as well as in vivo Ag-specific cytotoxic responses. Moreover, the fd-TSKB20 vaccine was able to protect against mortality induced by a high-dose inoculum of the parasite. Although vaccination with rfd phages successfully reduced both parasitemia and parasite load in the myocardium of WT B6 mice, Tlr9−/− animals were not protected against infection. Thus, our data extend previous studies, demonstrating that rfd phages induce Ag-specific IgG and CD8+ T cell-mediated responses and confer protection against an important human parasite infection, through a TLR9-dependent mechanism

Evaluation of Antibacterial Activity of Aminoglycosides and Modulating the Essential Oil of Cymbopogon citratus (DC.) Stapf

<p class="Standard" style="margin-bottom: .0001pt; text-align: justify; line-height: 200%;"> </p><div id=":tq.ma" class="Mu SP"><div id=":tq.co">Several works demonstrated the importance of the study of natural products as an alternative source for new antimicrobial drugs or for modulators for these ones. In this point, the aim of this was to investigate the antibacterial activity and the possible interactions between the essential oil of Cymbopogon citratus alone and in association with aminoglycosides against standard and clinically isolated strains of multidrug-resistant bacteria such as S. aureus, E. coli and P. aeruginosa by microdilution method. The results indicated a synergism between the antibiotics and the essential oil with a subinhibitory concentration (MIC/8), reducing the minimal inhibitory concentration (MIC) sixteen times against the multidrug-resistant strains of S. aureus 358, E. coli 27 and P. aeruginosa 143, but none modulatory activity was observed against P. aeruginosa 78 and P. aeruginosa 91 strains. By our results, can be concluded that the essential oil of Cymbopogon citratus can be an interesting source of natural products with antibacterial and/or modulatory antibiotic activitie</div></div><p class="Standard" style="margin-bottom: .0001pt; text-align: justify; line-height: 200%;"><span style="font-size: 12pt; line-height: 200%; font-family: 'Times New Roman', serif;"><span>AVALIAÇÃO DA ATIVIDADE ANTIBACTERIANA E MODULADORA DE AMINOGLICOSÍDEOS DO ÓLEO ESSENCIAL DE Cymbopogon citratus (DC.) STAPF</span></span></p><p class="Standard" style="margin-bottom: .0001pt; text-align: justify; line-height: 200%;"><span style="font-size: 12pt; line-height: 200%; font-family: 'Times New Roman', serif;"><span><span>Vários trabalhos vêm demonstrando a importância do estudo de produtos naturais como fonte alternativa para novos antimicrobianos ou que venham potencializar os já existentes. Neste contexto este trabalho teve como objetivo investigar a atividade antibacteriana e as possíveis interações entre o óleo essencial de Cymbopogon citratus combinados a aminoglicosídeos frente a linhagens padrões e multirresistentes de S. aureus, E. coli e de P. aeruginosa provenientes de isolados clínicos. Um ensaio de microdiluição foi realizado para verificar a atividade antibacteriana e as possíveis interacções entre o produto natural e os antibióticos, utilizando uma concentração sub-inibitória. Através dos resultados foi constatado a interferência sinérgica dos aminoglicosídeos quando associados com o óleo essencial em uma concentração de CIM/8, com redução das CIMs em até quatro pontos frente às linhagens de S. aureus 358, E. coli 27 e P. aeruginosa-143. Mas nenhuma atividade modificadora foi observada frente a P. aeruginosa 78 e P. aeruginosa 91. Através dos resultados pode-se concluir que o óleo essencial de Cymbopogon citratus pode ser uma fonte alternativa de produtos naturais com atividade antibacteriana.</span></span></span></p><p class="Standard" style="margin-bottom: .0001pt; text-align: justify; line-height: 200%;"><span style="font-size: 12pt; line-height: 200%; font-family: 'Times New Roman', serif;"><br /></span></p><p class="Standard" style="margin-bottom: .0001pt; text-align: justify; line-height: 200%;"><span style="font-size: 12pt; line-height: 200%; font-family: 'Times New Roman', serif;"><br /></span></p><p class="Standard" style="margin-bottom: .0001pt; text-align: justify; line-height: 200%;"><span style="font-size: 12pt; line-height: 200%; font-family: 'Times New Roman', serif;">Vários trabalhos vêm demonstrando a importância do estudo de produtos naturais como fonte alternativa para novos antimicrobianos ou que venham potencializar os já existentes. Neste contexto este trabalho teve como objetivo investigar a atividade antibacteriana e as possíveis interações entre o óleo essencial de <em>Cymbopogon citratus </em>combinados a aminoglicosídeos frente a linhagens padrões e multirresistentes de <em>S. aureus,</em> <em>E. coli </em>e de <em>P. aeruginosa </em>provenientes de<em> </em>isolados clínicos. Um ensaio de microdiluição foi </span><span style="font-size: 12pt; line-height: 200%; font-family: 'Times New Roman', serif;" lang="PT">realizado para verificar a atividade antibacteriana e as possíveis interacções entre o produto natural e os antibióticos, utilizando uma concentração sub-inibitória. Através  dos resultados foi constatado a </span><span style="font-size: 12pt; line-height: 200%; font-family: 'Times New Roman', serif;">interferência sinérgica  dos aminoglicosídeos quando associados com o óleo essencial em uma concentração de CIM/8, com redução das CIMs em  até quatro pontos frente  às linhagens de <em>S. aureus </em>358, <em>E. coli </em>27 e <em>P. aeruginosa-</em>143. Mas nenhuma atividade modificadora foi observada frente a <em>P. aeruginosa</em> 78 e <em>P. aeruginosa </em>91. Através dos resultados pode-se concluir que o óleo essencial de <em>Cymbopogon citratus </em>pode ser uma fonte alternativa de produtos naturais com atividade antibacteriana.</span></p

data_sheet_1_Vaccination With Recombinant Filamentous fd Phages Against Parasite Infection Requires TLR9 Expression.PDF

<p>Recombinant filamentous fd bacteriophages (rfd) expressing antigenic peptides were shown to induce cell-mediated immune responses in the absence of added adjuvant, being a promising delivery system for vaccination. Here, we tested the capacity of rfd phages to protect against infection with the human protozoan Trypanosoma cruzi, the etiologic agent of Chagas Disease. For this, C57BL/6 (B6) and Tlr9⁻^/⁻ mice were vaccinated with rfd phages expressing the OVA_257–264 peptide or the T. cruzi-immunodominant peptides PA8 and TSKB20 and challenged with either the T. cruzi Y-OVA or Y-strain, respectively. We found that vaccination with rfd phages induces anti-PA8 and anti-TSKB20 IgG production, expansion of Ag-specific IFN-γ, TNF-α, and Granzyme B-producing CD8⁺ T cells, as well as in vivo Ag-specific cytotoxic responses. Moreover, the fd-TSKB20 vaccine was able to protect against mortality induced by a high-dose inoculum of the parasite. Although vaccination with rfd phages successfully reduced both parasitemia and parasite load in the myocardium of WT B6 mice, Tlr9^−/− animals were not protected against infection. Thus, our data extend previous studies, demonstrating that rfd phages induce Ag-specific IgG and CD8⁺ T cell-mediated responses and confer protection against an important human parasite infection, through a TLR9-dependent mechanism.</p

Membrane Cholesterol Removal Changes Mechanical Properties of Cells and Induces Secretion of a Specific Pool of Lysosomes

<div><p>In a previous study we had shown that membrane cholesterol removal induced unregulated lysosomal exocytosis events leading to the depletion of lysosomes located at cell periphery. However, the mechanism by which cholesterol triggered these exocytic events had not been uncovered. In this study we investigated the importance of cholesterol in controlling mechanical properties of cells and its connection with lysosomal exocytosis. Tether extraction with optical tweezers and defocusing microscopy were used to assess cell dynamics in mouse fibroblasts. These assays showed that bending modulus and surface tension increased when cholesterol was extracted from fibroblasts plasma membrane upon incubation with MβCD, and that the membrane-cytoskeleton relaxation time increased at the beginning of MβCD treatment and decreased at the end. We also showed for the first time that the amplitude of membrane-cytoskeleton fluctuation decreased during cholesterol sequestration, showing that these cells become stiffer. These changes in membrane dynamics involved not only rearrangement of the actin cytoskeleton, but also <i>de novo</i> actin polymerization and stress fiber formation through Rho activation. We found that these mechanical changes observed after cholesterol sequestration were involved in triggering lysosomal exocytosis. Exocytosis occurred even in the absence of the lysosomal calcium sensor synaptotagmin VII, and was associated with actin polymerization induced by MβCD. Notably, exocytosis triggered by cholesterol removal led to the secretion of a unique population of lysosomes, different from the pool mobilized by actin depolymerizing drugs such as Latrunculin-A. These data support the existence of at least two different pools of lysosomes with different exocytosis dynamics, one of which is directly mobilized for plasma membrane fusion after cholesterol removal.</p></div

Cholesterol sequestration induces <i>de novo</i> actin polymerization.

<p>Fibroblasts were submitted to the following treatments: 95 nM of Latrunculin-A (Lat A) for one hour followed by washing with HBSS+ and incubation with fresh DMEM, without serum, for 45 minutes (A–C); previous incubation with Lat-A followed by washing with HBSS+ and incubation with 5 (D–F) or 10 (G–I) mM of MβCD diluted in fresh DMEM, without serum, for 45 minutes. The cells were fixed simultaneously and processed for F-actin cytoskeleton labeling using Phalloidin-FITC. Cell nuclei were stained with DAPI. Three independent experiments were performed in triplicates and at least 50 different cells were observed per group. Each selected image represents the most predominant actin cytoskeleton cell morphology of that experimental condition. Scale bars are all 10 µm.</p