A Proposed Mechanism of the Influence of Gold Nanoparticles on DNA Hybridization

A combination of gold nanoparticles (AuNPs) and nucleic acids has been used in biosensing applications. However, there is a poor fundamental understanding of how gold nanoparticle surfaces influence the DNA hybridization process. Here, we measured the rate constants of the hybridization and dehybridization of DNA on gold nanoparticle surfaces to enable the determination of activation parameters using transition state theory. We show that the target bases need to be detached from the gold nanoparticle surfaces before zipping. This causes a shift of the rate-limiting step of hybridization to the mismatch-sensitive zipping step. Furthermore, our results propose that the binding of gold nanoparticles to the single-stranded DNA segments (commonly known as bubbles) in the duplex DNA stabilizes the bubbles and accelerates the dehybridization process. We employ the proposed mechanism of DNA hybridization/dehybridization to explain the ability of 5 nm diameter gold nanoparticles to help discriminate between single base-pair mismatched DNA molecules when performed in a NanoBioArray chip. The mechanistic insight into the DNA–gold nanoparticle hybridization/dehybridization process should lead to the development of new biosensors

Dip-in Indicators for Visual Differentiation of Fuel Mixtures Based on Wettability of Fluoroalkylchlorosilane-Coated Inverse Opal Films

We have developed the dip-in indicator based on the inverse opal film (IOF) for visual differentiation of organic liquid mixtures, such as oil/gasoline or ethanol/gasoline fuel mixtures. The IOF consists of a three-dimensional porous structure with a highly ordered periodic arrangement of nanopores. The specularly reflected light at the interface of the nanopores and silica walls contributes to the structural color of the IOF film. This color disappears when the nanopores are infiltrated by a liquid with a similar refractive index to silica. The disappearance of the structural color provides a means to differentiate various liquid fuel mixtures based on their wettability of the nanopores in the IOF-based indicators. For differentiation of various liquid mixtures, we tune the wettability threshold of the indicator in such a way that it is wetted (color disappears) by one liquid but is not wetted by the other (color remains). Although colorimetric differentiation of liquids based on IOF wettability has been reported, differentiation of highly similar liquid mixtures require complicated readout approaches. It is known that the IOF wettability is controlled by multiple surface properties (e.g., oleophobicity) and structural properties (e.g., neck angle and film thickness) of the nanostructure. Therefore, we aim to exploit the combined tuning of these properties for differentiation of fuel mixtures with close compositions. In this study, we have demonstrated that, for the first time, the IOF-based dip-in indicator is able to detect a slight difference in the fuel mixture composition (i.e., 0.4% of oil content). Moreover, the color/no-color differentiation platform is simple, powerful, and easy-to-read. This platform makes the dip-in indicator a promising tool for authentication and determination of fuel composition at the point-of-purchase or point-of-use

Fotografia B221

The front-line treatment for adult acute myeloid leukemia (AML) is anthracycline-based combination chemotherapy. However, treatment outcomes remain suboptimal with relapses frequently observed. Among the mechanisms of treatment failure is multidrug resistance (MDR) mediated by the ABCB1, ABCC1, and ABCG2 drug-efflux transporters. Although genetic and phenotypic heterogeneity between leukemic blast cells is a well-recognized phenomenon, there remains minimal data on differences in MDR activity at the individual cell level. Specifically, functional assays that can distinguish the variability in MDR activity between individual leukemic blasts are lacking. Here, we outline a new dielectrophoretic (DEP) chip-based assay. This assay permits measurement of drug accumulation in single cells, termed same-single-cell analysis in the accumulation mode (<i>SASCA-A</i>). Initially, the assay was optimized in pretherapy samples from 20 adults with AML whose leukemic blasts had MDR activity against the anthracyline daunorubicin (DNR) tested using multiple MDR inhibitors. Parameters tested were initial drug accumulation, time to achieve signal saturation, fold-increase of DNR accumulation with MDR inhibition, ease of cell trapping, and ease of maintaining the trapped cells stationary. This enabled categorization into leukemic blast cells with MDR activity (MDR⁺) and leukemic blast cells without MDR activity (MDR^–ve). Leukemic blasts could also be distinguished from benign white blood cells (notably these also lacked MDR activity). MDR^–ve blasts were observed to be enriched in samples taken from patients who went on to enter complete remission (CR), whereas MDR⁺ blasts were frequently observed in patients who failed to achieve CR following front-line chemotherapy. However, pronounced variability in functional MDR activity between leukemic blasts was observed, with MDR⁺ cells not infrequently seen in some patients that went on to achieve CR. Next, we tested MDR activity in two paired AML patient samples. Pretherapy samples taken from patients that achieved CR to front-line chemotherapy were compared with samples taken at time of subsequent relapse. MDR⁺ cells were frequently observed in leukemic blast cells in both pretherapy and relapsed samples, consistent with MDR as a mechanism of relapse in these patients. We demonstrate the ability of a new DEP microfluidic chip-based assay to identify heterogeneity in MDR activity in leukemic blasts. The test provides a platform for future studies to characterize the mechanistic basis for heterogeneity in MDR activity at the individual cell level