RESUMO
The discovery of new alternatives for the treatment of infectious diseases has become the focus of burgeoning global interest. The complexation of the wide-spectrum antibiotic nalidixic acid (NA) with oxidovanadium(IV) ion and its incorporation into hybrid nanoparticulate systems were explored. The V-NA complex proved to be a stronger antimicrobial agent against E. coli, B. cereus, S. aureus and P. aeruginosa than NA, based on inhibition experiments. Myristyl myristate nanostructured lipid carriers (NLCs) and polymeric nanoparticles of Eudragit NE30D (EuNPs) were hybridized with chitosan (chi) to increase their stability and mucoadhesivity. They showed V-NA encapsulation of 97.8 ± 0.5% and 96.1 ± 0.1% respectively. TEM and DLS characterization ascertained the presence of spherical positive charged NPs ranging from 170 to 330 nm. Controlled release of V-NA from NPs was observed with 30-40% release in 3 days. A considerable potentiation of V-NA antimicrobial activity from 5 to 10 times was elucidated against P. aeruginosa with MIC values of 59.3 and 129.9 µM for NLC/chi and EuNPs/chi respectively, in comparison with 625 µM of the free complex. Hybrid NPs were able to interfere with the quorum sensing of the reporter Chromobacterium violaceum. Cytotoxicity on mouse fibroblast L929 cells was evaluated in the range of 29.7-519 µM by MTT assay showing that, NLC/chi particles supported cell growth in the range of at 29.7-60 µM while Eu/chi do not exert cytotoxicity between 29.7 and 120 µM. These results suggest that nanoparticles are suitable systems for drug delivery applications.
Assuntos
Antibacterianos/química , Antibacterianos/farmacologia , Quitosana/química , Complexos de Coordenação/química , Nanopartículas Metálicas/química , Ácido Nalidíxico/química , Percepção de Quorum/efeitos dos fármacos , Vanádio/química , Animais , Linhagem Celular , Sobrevivência Celular , Portadores de Fármacos/química , Resistencia a Medicamentos Antineoplásicos , Camundongos , Tamanho da PartículaRESUMO
Medical cotton gauzes were modified by grafting poly(methacrylic acid) (PMAA) via free radical polymerization to obtain wound dressings with antimicrobial and drug delivery properties. The effect of several reaction parameters including monomer and initiator concentrations, reaction time, and temperature was studied. The grafting was confirmed by Fourier transform infrared spectroscopy (FT-IR), thermogravimetric analysis (TGA), swelling studies, and scanning electron microscopy (SEM). The grafted cotton gauzes (gauze-g-PMAA) samples were loaded with ZnO nanoparticles to endow with antibacterial properties. Also, they were tested as drug eluting systems using nalidixic acid as antimicrobial agent. The antibacterial activity of the ZnO-loaded gauze-g-PMAA samples was evaluated against Escherichia coli (E. coli) and Staphylococcus epidermidis (S. epidermidis). The PMAA-grafted gauzes showed antibacterial activity and inhibited the growth of both microorganisms. These results suggest that the PMAA-grafted cotton gauzes could be used in the biomedical area particularly as antimicrobial and drug-eluting wound dressings.
Assuntos
Antibacterianos/química , Bandagens , Celulose/química , Fibra de Algodão , Sistemas de Liberação de Medicamentos , Ácidos Polimetacrílicos/química , Celulose/síntese química , Liberação Controlada de Fármacos , Escherichia coli/efeitos dos fármacos , Ácido Nalidíxico/química , Ácido Nalidíxico/farmacologia , Nanopartículas/química , Polimerização , Ácidos Polimetacrílicos/síntese química , Staphylococcus epidermidis/efeitos dos fármacos , Óxido de Zinco/química , Óxido de Zinco/farmacologiaRESUMO
OBJECTIVES: The aim of this study was to characterise OXA-258 variants and other features that may contribute to carbapenem resistance in Achromobacter ruhlandii. METHODS: Kinetic parameters for purified OXA-258a and OXA-258b were determined measuring the rate of hydrolysis of a representative group of antimicrobial agents. Whole-genome shotgun sequencing was performed on A. ruhlandii 38 (producing OXA-258a) and A. ruhlandii 319 (producing OXA-258b), and in silico analysis of antimicrobial resistance determinants was conducted. Substrates of the AxyABM efflux pump were investigated by inhibition assays using phenylalanine-arginine ß-naphthylamide (PAßN). Outer membrane protein profiles were resolved by 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). RESULTS: Kinetic measurements of purified OXA-258 variants displayed an overall weak catalytic efficiency toward ß-lactams. A detectable hydrolysis of imipenem was observed. In silico genomic analysis confirmed the presence of 32 and 35 putative efflux pump-encoding genes in A. ruhlandii strains 38 and 319, respectively. Complete sequences for AxyABM and AxyXY efflux pumps, previously described in Achromobacter xylosoxidans, were detected. Decreases in the MICs for chloramphenicol, nalidixic acid and trimethoprim/sulfamethoxazole were observed in the presence of the inhibitor PAßN, suggesting that these antibiotics are substrates of AxyABM. AxyXY-encoding genes of A. ruhlandii 38 and A. ruhlandii 319 displayed 99% identity. No differences were observed in the outer membrane protein profiles. CONCLUSIONS: The contribution of OXA-258 enzymes to the final ß-lactam resistance profile may be secondary. Further studies on other putative resistance markers identified in the whole-genome analysis should be conducted to understand the carbapenem resistance observed in A. ruhlandii.
Assuntos
Achromobacter/enzimologia , Antibacterianos/farmacologia , Proteínas da Membrana Bacteriana Externa/genética , Sequenciamento Completo do Genoma/métodos , Resistência beta-Lactâmica , beta-Lactamases/genética , Achromobacter/genética , Antibacterianos/química , Proteínas de Bactérias/genética , Cloranfenicol/química , Cloranfenicol/farmacologia , Simulação por Computador , Variação Genética , Hidrólise , Imipenem/química , Imipenem/farmacologia , Testes de Sensibilidade Microbiana , Ácido Nalidíxico/química , Ácido Nalidíxico/farmacologia , Combinação Trimetoprima e Sulfametoxazol/química , Combinação Trimetoprima e Sulfametoxazol/farmacologiaRESUMO
We have synthesized two naphthyl ester quinolone derivates and determined their ability to generate reactive oxygen species (ROS) such as (1)O(2), ()OH, H(2)O(2) upon photolysis with UV-A light. The ability of cinoxacin (1) and nalidixic acid (2), and their naphthyl ester derivatives (3 and 4) to generate a dose-dependent amount of singlet oxygen and ROS (()(-)O(2), ()OH) in cell-free systems was detected by histidine assay and by luminol-enhanced chemiluminescence (LCL), respectively. Their electronic absorption and emission spectra were quantified and their photostability was determined. Their tendency to generate peroxidic derivative species showed the following order: 3>4; in contrast, their ability to generate singlet oxygen was 4>3 and these were better sensitizers than their parent quinolones 1 and 2. The antibacterial activity in darkness and under irradiation of compounds 3 and 4 was tested on Escherichia coli and compared with that of their parent compounds. An enhanced antibacterial activity by irradiation of the naphthyl esters of cinoxacin and nalidixic acid on E. coli was observed.
Assuntos
Antibacterianos/química , Cinoxacino/química , Ácido Nalidíxico/química , Naftalenos/química , Antibacterianos/síntese química , Antibacterianos/farmacologia , Sobrevivência Celular , Cinoxacino/síntese química , Cinoxacino/farmacologia , Escherichia coli/efeitos dos fármacos , Escherichia coli/metabolismo , Radical Hidroxila/química , Radical Hidroxila/metabolismo , Ácido Nalidíxico/síntese química , Ácido Nalidíxico/farmacologia , Naftalenos/síntese química , Naftalenos/farmacologia , Naftiridinas , Oxigênio/química , Oxigênio/metabolismo , Espécies Reativas de Oxigênio/química , Espécies Reativas de Oxigênio/metabolismoRESUMO
The photophysics and photochemistry of nalidixic acid (NA) were studied as function of pH and solvent properties. The ground state of NA exhibits different protonated forms in the range of pH 1.8-10.0. Fluorescence studies showed that the same species exist at the lowest singlet excited state. Absorption experiments were carried out with NA and with the methylated analog of nalidixic acid (MNE) in different organic solvents and water pH 3, where the main species corresponds to that protonated at the carboxylic group. These studies and the DFT calculation of torsional potential energy profiles suggest that the most stable conformation of the NA in nonprotic solvents corresponds to a closed structure caused by the existence of intramolecular hydrogen bond. Absorption and fluorescence spectra were studied in sulfuric acid solution. The pK value (Ho -1.0) found in these conditions was attributed to the protonation of the 4' keto oxygen atom of the heterocyclic ring. Theoretical calculations (DFT/B3LYP/6-311G*) of the energies of the different monoprotonated forms of the NA and Fukui indexes (f(x)-) showed that the species with the proton attached to 4' keto oxygen atom is the most stable of all the cationic forms. MNE and enoxacin also showed the protonation of the 4' keto oxygen atom with similar pK values. The photodecomposition of NA is dependent on the medium properties. Faster decomposition rates were obtained in strong acid solution. In nonprotic solvents, a very slow decomposition rate was observed.