Anaerobic activation of benzene is expected to represent a novel biochemistry of environmental significance. Therefore, benzene metabolism was investigated inGeobacter metallireducens, the only genetically tractable organism known to anaerobically degrade benzene. Trace amounts (<0.5 μM) of phenol accumulated in cultures ofGeobacter metallireducensanaerobically oxidizing benzene to carbon dioxide with the reduction of Fe(III). Phenol was not detected in cell-free controls or in Fe(II)- and benzene-containing cultures ofGeobacter sulfurreducens, aGeobacterspecies that cannot metabolize benzene. The phenol produced inG. metallireducenscultures was labeled with18O during growth in H218O, as expected for anaerobic conversion of benzene to phenol. Analysis of whole-genome gene expression patterns indicated that genes for phenol metabolism were upregulated during growth on benzene but that genes for benzoate or toluene metabolism were not, further suggesting that phenol was an intermediate in benzene metabolism. Deletion of the genes for PpsA or PpcB, subunits of two enzymes specifically required for the metabolism of phenol, removed the capacity for benzene metabolism. These results demonstrate that benzene hydroxylation to phenol is an alternative to carboxylation for anaerobic benzene activation and suggest that this may be an important metabolic route for benzene removal in petroleum-contaminated groundwaters, in whichGeobacterspecies are considered to play an important role in anaerobic benzene degradation.
Cyanobacteria constitute a morphologically diverse group of oxygenic photoautotrophic microbes which range from unicellular to multicellular, and non-nitrogen-fixing to nitrogen-fixing types. Sustained long-term exposure to changing environmental conditions, during their three billion years of evolution, has presumably led to their adaptation to diverse ecological niches. The ability to maintain protein conformational homeostasis (folding–misfolding–refolding or aggregation–degradation) by molecular chaperones holds the key to the stress adaptability of cyanobacteria. Although cyanobacteria possess several genes encoding DnaK and DnaJ family proteins, these are not the most abundant heat-shock proteins (Hsps), as is the case in other bacteria. Instead, the Hsp60 family of proteins, comprising two phylogenetically conserved proteins, and small Hsps are more abundant during heat stress. The contribution of the Hsp100 (ClpB) family of proteins and of small Hsps in the unicellular cyanobacteria (SynechocystisandSynechococcus) as well as that of Hsp60 proteins in the filamentous cyanobacteria (Anabaena) to thermotolerance has been elucidated. The regulation of chaperone genes by severalcis-elements andtrans-acting factors has also been well documented. Recent studies have demonstrated novel transcriptional and translational (mRNA secondary structure) regulatory mechanisms in unicellular cyanobacteria. This article provides an insight into the heat-shock response: its organization, and ecophysiological regulation and role of molecular chaperones, in unicellular and filamentous nitrogen-fixing cyanobacterial strains.
Surface functionalization, colloidal stability and biocompatibility of magnetic nanoparticles are crucial for their biological applications. Here, we report a synthetic approach for the direct preparation of superparamagnetic nanoparticles consisting of a perovskite LSMO core modified with a covalently linked chitosan shell that provides colloidal stability in aqueous solutions for cancer hyperthermia therapy. The characterization of the core-shell nanostructure using Fourier transform infrared spectroscopy; thermo-gravimetric analysis to assess the chemical bonding of chitosan to nanoparticles; field-emission scanning electron microscopy and transmission electron microscopy for its size and coating efficiency estimation; and magnetic measurement for their magnetization properties was performed. Zeta potential and light scattering studies of the core shell revealed it to possess good colloidal stability. Confocal microscopy and MTT assay are performed for qualitative and quantitative measurement of cell viability and biocompatibility. In depth cell morphology and biocompatibility is evaluated by using multiple-staining of different dyes. The magnetic@chitosan nanostructure system is found to be biocompatible up to 48 h with 80% cell viability. Finally, an in vitro cancer hyperthermia study is done on the MCF7 cell line. During in vitro hyperthermia treatment of cancer cells, cell viability is reduced upto 40% within 120 min with chitosan coated nanoparticles. Our results demonstrate that this simplified and facile synthesis strategy shows potential for designing a colloidal stable state and biocompatible core shell nanostructures for cancer hyperthermia therapy.
Antibiotic resistant bacteria not only affect human health and but also threatens the safety in hospitals and among communities. However, the emergence of drug resistant bacteria is inevitable due to evolutionary selection as a consequence of indiscriminate antibiotic usage. Therefore, it is necessary to develop a novel strategy by which pathogenic bacteria can be eliminated without triggering resistance. We propose a novel magnetic nanoparticle-based physical treatment against pathogenic bacteria, which blocks biofilm formation and kills bacteria. In this approach, multiple drug resistant Staphylococcus aureus USA300 and uropathogenic Escherichia coli CFT073 are trapped to the positively charged magnetic core-shell nanoparticles (MCSNPs) by electrostatic interaction. All the trapped bacteria can be completely killed within 30 min owing to the loss of membrane potential and dysfunction of membrane-associated complexes when exposed to the radiofrequency current. These results indicate that MCSNP-based physical treatment can be an alternative antibacterial strategy without leading to antibiotic resistance, and can be used for many purposes including environmental and therapeutic applications.