Recombinant Escherichia coli (E. coli) bacteria expressing green fluorescent protein (GFP) was used as a model system to investigate the antimicrobial activities of Ag nanoparticles (NPs). A convenient in situ method of Ag NP synthesis using sodium borohydride, in the bacterial growth medium, was developed to produce preformed NPs for the study. Fluorescence spectroscopic and microscopic techniques allowed rapid detection of time-dependent changes in bacterial growth as well as fluorescence characteristics in the presence of Ag NPs. In addition, X-ray diffraction, UV-vis spectroscopic, and transmission electron microscopic measurements were carried out to understand the effect of Ag NPs on the bacteria. Our observations indicated that Ag NPs, above a certain concentration, not only were bactericidal but also were found to reduce the sizes of treated bacteria in comparison to untreated ones. Cell lysis of Ag NP-treated bacteria was suggested by the increased GFP fluorescence obtained in the medium. In vitro DNA-Ag NP interaction was detected by spectrophotometric analysis. However, electrophoresis studies indicated no direct effect of Ag NPs on DNA or protein profiles.
Highly fluorescent copper nanoclusters (Cu NCs) have been synthesized using single-step reduction of copper sulfate by hydrazine in the presence of lysozyme. The fluorescence quantum yield was measured to be as high as 18%. The emission was also found to be dependent on the excitation wavelength. Mass spectrometric analyses indicated the presence of species corresponding to Cu2 to Cu9. Transmission electron microscopic analyses indicated the formation of agglomerated particles of average diameter of 2.3 nm, which were constituted of smaller particles of average diameter of 0.96 nm. They were found to be stable between pH 4 and 10 and in addition having excellent chemical and photostability. The noncytotoxic NCs were used to successfully label cervical cancer HeLa cells.
We report the finding of the presence of carbon nanoparticles (CNPs) in different carbohydrate based food caramels, viz. bread, jaggery, sugar caramel, corn flakes and biscuits, where the preparation involves heating of the starting material. The CNPs were amorphous in nature; the particles were spherical having sizes in the range of 4–30 nm, depending upon the source of extraction. The results also indicated that particles formed at higher temperature were smaller than those formed at lower temperature. Excitation tuneable photoluminescence was observed for all the samples with quantum yield (QY) 1.2, 0.55 and 0.63%, for CNPs from bread, jaggery and sugar caramels respectively. The present discovery suggests potential usefulness of CNPs for various biological applications, as the sources of extraction are regular food items, some of which have been consumed by humans for centuries, and thus they can be considered as safe.
Herein we report synergy in antimicrobial activity of a chitosan-silver nanoparticle (CS-Ag NP) composite in the presence of molecular iodine. Green fluorescent protein (GFP) expressing recombinant Escherichia coli bacteria have been used to test the efficacy and establish the mechanism of action. Experimental evidence indicate significantly high bactericidal activity of the nanocomposite in the presence of iodine than either due to the composite, chitosan, Ag NP or iodine only. Transmission electron microscopy measurements revealed attachment of bacteria to the composite. In addition, flow cytometry results supported definite occurrence of cell wall damage of the bacteria treated with the composite in the presence of iodine. Further, the nanocomposite and iodine combination was found to exert reactive oxygen species (ROS) generated oxidative stress in the cytoplasm of bacterial cells, leading to cell death. Elucidation of the mechanism of synergy due to three potential antibacterial components suggested that on the surface of Ag NPs molecular iodine possibly generated iodine atom thus contributing toward free radical induced oxidative stress, whereas chitosan and Ag NPs facilitated the process of cell killing and thus collectively enhanced the potency of antimicrobial effect at the lowest concentrations of individual components.
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