RESUMEN
Oral cancer is one of the most common types of cancer in Europe and its large diffusion requires, together with prevention, the development of low-cost and reliable portable platforms for its diagnosis, with features of high selectivity and sensitivity. In this study, the development and characterization of a molecularly imprinted polymer (MIP)-based electrochemical sensor for TGF-ß1 detection are reported. The optimized biosensor is a potential tool for the early screening of oral cancer. A biomimetic surface has been obtained by electropolymerization of o-phenylenediamine (o-PD) on platinum electrodes, in the presence of TGF-ß1 as a template molecule. MIP synthesis, template removal and TGF-ß1 rebinding have been monitored by Differential Pulse Voltammetry (DPV). Atomic Force Microscopy (AFM) has been performed to investigate and characterize the surface morphology and the influence of the washing step on MIP and NIP (non-imprinted polymer as the control) while the thickness of the polymer layer has been measured by Scanning Transmission Electron Microscopy (STEM) analysis. The MIP sensor performance has been tested in both buffer solution and saliva samples with TGF-ß1, showing a linear response in the considered range (from 20 ng ml-1 down to 0.5 ng ml-1), an outstanding LOD of 0.09 ng mL-1 and affinity and selectivity to TGF-ß1 also in the presence of interfering molecules. The sensor was used also for the detection of target molecules in spiked saliva samples with good recovery results suggesting the possibility of the use of the proposed system for large scale fast screening in oral cancer diagnosis.
Asunto(s)
Polímeros Impresos Molecularmente , Neoplasias de la Boca , Humanos , Factor de Crecimiento Transformador beta1 , Neoplasias de la Boca/diagnóstico , Polímeros , Biopsia LíquidaRESUMEN
Pillar-like structures of nanodiamonds on a silicon substrate are self-assembled for the first time by a pulsed spray technique. This technique allows us to deposit nanodiamond layers by using high quality nanocrystals of 250 nm dispersed in 1,2-dichloroethane (DCE) solvent. The analysis of 2D/3D confocal and atomic force microscopy images evidences the presence of self-assembled pillar-like structures distributed in an irregular way. The proposed method is simple, easy and cheap, and does not require complex growth processes or structured materials, ideal for upscaling toward industrial biochip implementation and photonic applications. The suggested formation mechanisms of self-assembly are based on the so-called coffee stain effect, i.e., on the time evolution of DCE evaporation.
RESUMEN
In this study, the evidence of electron-dense magnetic inclusions with polyhedral shape in the cytoplasm of Harveyi clade Vibrio strain PS1, a bioluminescent bacterium living in symbiosis with marine organisms, led us to investigate the behavior of this bacterium under exposure to static magnetic fields ranging between 20 and 2000 Gauss. When compared to sham-exposed, the light emission of magnetic field-exposed bacteria growing on solid medium at 18°C ±0.1°C was increased up to two-fold as a function of dose and growth phase. Stimulation of bioluminescence by magnetic field was more pronounced during the post-exponential growth and stationary phase, and was lost when bacteria were grown in the presence of the iron chelator deferoxamine, which caused disassembly of the magnetic inclusions suggesting their involvement in magnetic response. As in luminescent Vibrio spp. bioluminescence is regulated by quorum sensing, possible effects of magnetic field exposure on quorum sensing were investigated. Measurement of mRNA levels by reverse transcriptase real time-PCR demonstrated that luxR regulatory gene and luxCDABE operon coding for luciferase and fatty acid reductase complex were significantly up-regulated in magnetic field-exposed bacteria. In contrast, genes coding for a type III secretion system, whose expression was negatively affected by LuxR, were down-regulated. Up-regulation of luxR paralleled with down-regulation of small RNAs that mediate destabilization of luxR mRNA in quorum sensing signaling pathways. The results of experiments with the well-studied Vibrio campbellii strain BB120 (originally classified as Vibrio harveyi) and derivative mutants unable to synthesize autoinducers suggest that the effects of magnetic fields on quorum sensing may be mediated by AI-2, the interspecies quorum sensing signal molecule.
Asunto(s)
Campos Magnéticos , Percepción de Quorum , Vibrio/fisiología , Genes Bacterianos , Mediciones Luminiscentes , Mutación , Vibrio/genética , Vibrio/metabolismoAsunto(s)
Antifúngicos/uso terapéutico , Claritromicina/uso terapéutico , Fungemia/tratamiento farmacológico , Geotricosis/tratamiento farmacológico , Geotrichum , Antibacterianos/uso terapéutico , Fungemia/microbiología , Geotricosis/microbiología , Humanos , Masculino , Pruebas de Sensibilidad Microbiana , Persona de Mediana Edad , Infecciones por Pseudomonas/complicaciones , Infecciones por Pseudomonas/tratamiento farmacológico , Infecciones por Pseudomonas/microbiología , Sobreinfección/tratamiento farmacológico , Sobreinfección/microbiologíaRESUMEN
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