RESUMO
Pseudomonas aeruginosa (PA) is an important opportunistic pathogen that causes different infections on immunocompromised patients. Within PA accessory genome, differences in virulence, antibiotic resistance and biofilm formation have been described between strains, leading to the emergence of multidrug-resistant strains. The genome sequences of 17 strains isolated from patients with healthcare-associated infections in a Mexican hospital were genomically and phylogenetically analyzed and antibiotic resistance genes, virulence genes, and biofilm formation genes were detected. Fifteen of the 17 strains were resistant to at least two of the carbapenems meropenem, imipenem, and the monobactam aztreonam. The antibiotic resistance (mexA, mexB, and oprM) and the biofilm formation (pslA and pslD) genes were detected in all strains. Differences were found between strains in accessory genome size. The strains had different sequence types, and seven strains had sequence types associated with global high risk epidemic PA clones. All strains were represented in two groups among PA global strains. In the 17 strains, horizontally acquired resistance genes to aminoglycosides and beta-lactams were found, mainly, and between 230 and 240 genes that encode virulence factors. The strains under study were variable in terms of their accessory genome, antibiotic resistance, and virulence genes. With these characteristics, we provide information about the genomic diversity of clinically relevant PA strains.
Assuntos
Carbapenêmicos , Infecções por Pseudomonas , Humanos , Aztreonam , Pseudomonas aeruginosa/genética , Antibacterianos , Hospitais , Genômica , Atenção à Saúde , Testes de Sensibilidade MicrobianaRESUMO
Pseudomonas aeruginosa (PA) is considered the first causal agent of morbidity and mortality in people with cystic fibrosis (CF) disease. Multi-resistant strains have emerged due to prolonged treatment with specific antibiotics, so new alternatives have been sought for their control. In this context, there is a renewed interest in therapies based on bacteriophages (phages) supported by several studies suggesting that therapy based on lytic phages and biofilm degraders may be promising for the treatment of lung infections in CF patients. However, there is little clinical data about phage studies in CF and the effectiveness and safety in patients with this disease has not been clear. Therefore, studies regarding on phage characterization, selection, and evaluation in vitro and in vivo models will provide reliable information for designing effective cocktails, either using mixed phages or in combination with antibiotics, making a great progress in clinical research. Hence, this review focuses on the most relevant and recent findings on the activity of lytic phages against PA strains isolated from CF patients and hospital environments, and discusses perspectives on the use of phage therapy on the treatment of PA in CF patients.
Assuntos
Bacteriófagos , Fibrose Cística , Infecções por Pseudomonas , Fagos de Pseudomonas , Humanos , Pseudomonas aeruginosa , AntibacterianosRESUMO
Foodborne diseases are extremely relevant and constitute an area of alert for public health authorities due to the high impact and number of people affected each year. The food industry has implemented microbiological control plans that ensure the quality and safety of its products; however, due to the high prevalence of foodborne diseases, the industry requires new microbiological control systems. One of the main causative agents of diseases transmitted by poultry meat is the bacterium Salmonella enterica. Disinfectants, antibiotics, and vaccines are used to control this pathogen. However, they have not been efficient in the total elimination of these bacteria, with numerous outbreaks caused by this bacterium observed today, in addition to the increase in antibiotic-resistant bacteria. The search for new technologies to reduce microbial contamination in the poultry industry continues to be a necessity and the use of lytic bacteriophages is one of the new solutions. In this study, 20 bacteriophages were isolated for Salmonella spp. obtained from natural environments and cocktails composed of five of them were designed, where three belonged to the Siphoviridae family and two to the Microviridae family. This cocktail was tested on chicken meat infected with Salmonella Typhimurium at 10 °C, where it was found that this cocktail was capable of decreasing 1.4 logarithmic units at 48 h compared to the control.
RESUMO
The human red blood cell (RBC) membrane has significant elastic capabilities which can be described measuring typical membrane edge fluctuations and mechanical properties by optical techniques. The RBC elastic properties can be affected by changes in the surrounding media. In an attempt to elucidate the molecular mechanisms of the interaction of resveratrol with the red cell membrane and of its antioxidant capacity the changes in mechanical properties of the RBC membrane were analyzed. These studies were carried out through measurements of RBC membrane fluctuations in the presence of the oxidant agent HClO using thermal fluctuation spectroscopy (TFS). The observed results showed that the elastic capabilities of RBC changed with low concentration of hypochlorous acid but without morphological changes. However, in the presence of resveratrol the deformation and decrease of elastic capabilities induced by HClO on RBC decreased. These in vitro results demonstrated the protective effect of RV against the detrimental effects triggered by HClO upon human erythrocytes.
Assuntos
Antioxidantes/metabolismo , Eritrócitos/metabolismo , Resveratrol/sangue , Análise Espectral/métodos , Membrana Eritrocítica/metabolismo , Humanos , Ácido Hipocloroso/metabolismo , Análise de Célula ÚnicaRESUMO
Thimerosal (THI, ethyl-mercury thiosalicylate) is added to vaccines as a preservative; as a consequence, infants may have been exposed to bolus doses of Hg that collectively added up to nominally 200 µg Hg during the first 6 months of life. While several studies report an association between THI-containing vaccines and neurological disorders, other studies do not support the causal relation between THI and autism. With the purpose to understand the molecular mechanisms of the toxic effect of THI it was assayed on human red cells and in bilayers built-up of dimyristoylphosphatidylcholine (DMPC) and dimyristoylphosphatidylethanolamine (DMPE), classes of phospholipids found in the outer and inner monolayers of the human erythrocyte membrane, respectively. The capacity of THI to interact with DMPC and DMPE was determined by X-ray diffraction and differential scanning calorimetry, whereas intact human erythrocytes were observed by optical, defocusing and scanning electron microscopy. The experimental findings of this study demonstrated that THI interacted in a concentration-dependent manner with DMPC and DMPE bilayers, and in vitro interacted with erythrocytes inducing morphological changes. However, concentrations were considerable higher than those present in vaccines.
Assuntos
Eritrócitos/efeitos dos fármacos , Bicamadas Lipídicas , Timerosal/farmacologia , Varredura Diferencial de Calorimetria , Células Cultivadas , Dimiristoilfosfatidilcolina/química , Membrana Eritrocítica/química , Membrana Eritrocítica/efeitos dos fármacos , Eritrócitos/ultraestrutura , Humanos , Bicamadas Lipídicas/química , Lipossomos , Estrutura Molecular , Fosfatidiletanolaminas/química , Conservantes Farmacêuticos/farmacologia , Termodinâmica , Timerosal/química , Difração de Raios XRESUMO
Gallic acid (GA) is a polyphenol present in many plants. This study was aimed to investigate the molecular interaction of GA with the human erythrocyte membrane and to determine its antioxidant capacity. The molecular interaction with the membrane of human red cells and the antioxidant property was assayed on both human red cells and molecular models of its membrane. Observations by optical, scanning electron, and defocusing microscopy demonstrated that GA is capable to convert red cells from their normal biconcave shape to crenated echinocytes. This result indicates that GA molecules are positioned in the outer monolayer of the red cell membrane. Dimyristoylphosphatidylcholine (DMPC) and dimyristoylphosphatidylethanolamine (DMPE) were selected as classes of phospholipids found in the outer and inner monolayers of the red cell membrane, respectively. X-ray diffraction and differential scanning calorimetry showed that GA was preferentially bound to DMPC bilayers. Experiments related to the antioxidant capacity of GA indicated that this compound offsets HClO oxidative capacity on DMPE bilayers. In addition, optical, scanning, defocusing microscopy, and hemolysis assays confirmed the protective capacity of GA against HClO deleterious effects on human red cells. As a conclusion, GA would be capable to block the access of oxidants into the lipid bilayer, and thus avoid their access into red cells.
Assuntos
Antioxidantes/farmacologia , Eritrócitos/efeitos dos fármacos , Eritrócitos/metabolismo , Ácido Gálico/farmacologia , Antioxidantes/química , Varredura Diferencial de Calorimetria , Células Cultivadas , Membrana Eritrocítica/efeitos dos fármacos , Membrana Eritrocítica/metabolismo , Eritrócitos/ultraestrutura , Ácido Gálico/química , Hemólise/efeitos dos fármacos , Humanos , Bicamadas Lipídicas , Estrutura Molecular , Fosfolipídeos , Termodinâmica , Difração de Raios XRESUMO
In order to gain insight into the molecular mechanism of the antioxidant properties of Solanum crispum, aqueous extracts of its leaves were assayed on human erythrocytes and molecular models of its membrane. Phenolics and alkaloids were detected by HPLC-MS. Scanning electron and defocusing microscopy showed that S. crispum changed erythrocytes from the normal shape to echinocytes. These results imply that molecules present in the aqueous extracts were located in the outer monolayer of the erythrocyte membrane. Dimyristoylphosphatidylcholine (DMPC) and dimyristoylphosphatidylethanolamine (DMPE) were chosen as representative of phospholipid classes located in the outer and inner monolayers of the erythrocyte membrane, respectively. X-ray diffraction showed that S. crispum preferentially interacted with DMPC bilayers. Experiments regarding its antioxidant properties showed that S. crispum neutralized the oxidative capacity of HClO on DMPE bilayers; defocusing microscopy and hemolysis assays demonstrated the protective effect of S. crispum against the oxidant effects of HClO on human erythrocytes.
Assuntos
Antioxidantes/farmacologia , Eritrócitos/efeitos dos fármacos , Eritrócitos/metabolismo , Extratos Vegetais/farmacologia , Solanum/química , Cromatografia Líquida de Alta Pressão , Eritrócitos/citologia , Eritrócitos/ultraestrutura , Hemólise/efeitos dos fármacos , Humanos , Bicamadas Lipídicas , Espectrometria de Massas , Extratos Vegetais/química , Polifenóis/química , Polifenóis/farmacologia , Difração de Raios XRESUMO
Toxicity of vanadium on cells is one of the less studied effects. This prompted us to study the structural effects induced on neuroblastoma and erythrocytes by vanadium (V) sodium metavanadate. This salt was incubated with mice cholinergic neuroblastoma cells and intact human erythrocytes. To learn whether metavanadate interacts with membrane lipid bilayers it was incubated with bilayers built-up of dimyristoylphosphatidylcholine (DMPC) and dimyristoylphosphatidylethanolamine (DMPE). These are phospholipid classes located in the outer and inner monolayers of the human erythrocyte membrane, respectively. Exposure of neuroblastoma cells to metavanadate showed significant decreases in cell viability as well as in cell number correlating with inhibition of aconitase activity. In scanning electron microscopy (SEM) and defocusing microscopy (DM) it was observed that induced on erythrocytes the formation of echinocytes. However, no effects were obtained when metavanadate was made to interact with DMPC and DMPE multibilayers and liposomes, assays performed by X-ray diffraction and differential scanning calorimetry (DSC), respectively. These results imply that the effects of metavanadate on erythrocytes are through interactions with proteins located in the membrane outer moiety, and could still involve other minor lipid components as well. Also, partly unsaturated lipids could interact differently the fully saturated chains in the model systems.
Assuntos
Eritrócitos/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Vanadatos/farmacologia , Acetilcoenzima A/metabolismo , Animais , Varredura Diferencial de Calorimetria , Diferenciação Celular , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Dimiristoilfosfatidilcolina/química , Eritrócitos/citologia , Humanos , Lipossomos , Camundongos , Microscopia Eletrônica de Varredura , Neuroblastoma , Neurônios/citologia , Neurônios/metabolismo , Fosfatidiletanolaminas/química , Vanadatos/química , Vanadatos/toxicidade , Difração de Raios XRESUMO
We present a real-time method to measure the amplitude of thermal fluctuations in biological membranes by means of a new treatment of the defocusing microscopy (DM) optical technique. This approach was also applied to study the deformation of human erythrocytes to its echinocyte structure. This was carried out by making three-dimensional shape reconstructions of the cell and measuring the thermal fluctuations of its membrane, as the cell is exposed to the anti-inflammatory drug naproxen and as it recovers its original shape, when it is subsequently cleansed of the drug. The results showed biomechanical changes in the membrane even at low naproxen concentration (0.2 mM). Also, we found that when the cell recovered its original shape, the membrane properties were different compared to the nondrugged initial erythrocyte, indicating that the drug administration-recovery process is not completely reversible.