Peroxidase immobilized on phospholipid bilayers supported on au (111) by DTT self‐assembled monolayers: Application to dopamine determination

In this work, horseradish peroxidase (HRP) was immobilized on dimyristoylphosphatidylcholine (DMPC) bilayers supported on Au (111) by dithiotreitol (DTT) self‐assembled monolayers and used as a nanostructured electrochemical biosensor to dopamine determination. The morphology of the phospholipid bilayers and the immobilization of HRP to these layers were characterized by atomic force microscopy (AFM). Square‐wave voltammetry (SWV) experiments were done to investigate the performance of the HRP‐modified electrode. The AFM images indicate that the enzyme is adsorbed at the external layer of the lipid bilayer and, although the electrical charges on the surface were not measured, the enzyme and phospholipids surface interaction occurs probably by electrostatic forces due to the pH used in the experiments. Interestingly, the present system can be used as one‐shot sensor for the rapid detection of dopamine. The analytical performance of this system was linear for dopamine concentrations from 3.3 × 10^?5 to 1.3 × 10^?3 mol L^?1 (r = 0.9997) with a detection limit of 2.0 × 10^?6 mol L^?1. Our results indicate that the use of HRP‐DMPC bilayer system may be useful not only in developing new nanostructured materials for technological purposes, but could be very useful in fundamental studies to investigate the interactions between different micro‐and macromolecules, even with soluble proteins, and lipid membranes. Biotechnol. Bioeng. 2013; 110: 374–382. © 2012 Wiley Periodicals, Inc.     

Implementing vertex dynamics models of cell populations in biology within a consistent computational framework

The dynamic behaviour of epithelial cell sheets plays a central role during development, growth, disease and wound healing. These processes occur as a result of cell adhesion, migration, division, differentiation and death, and involve multiple processes acting at the cellular and molecular level. Computational models offer a useful means by which to investigate and test hypotheses about these processes, and have played a key role in the study of cell–cell interactions. However, the necessarily complex nature of such models means that it is difficult to make accurate comparison between different models, since it is often impossible to distinguish between differences in behaviour that are due to the underlying model assumptions, and those due to differences in the in silico implementation of the model. In this work, an approach is described for the implementation of vertex dynamics models, a discrete approach that represents each cell by a polygon (or polyhedron) whose vertices may move in response to forces. The implementation is undertaken in a consistent manner within a single open source computational framework, Chaste, which comprises fully tested, industrial-grade software that has been developed using an agile approach. This framework allows one to easily change assumptions regarding force generation and cell rearrangement processes within these models. The versatility and generality of this framework is illustrated using a number of biological examples. In each case we provide full details of all technical aspects of our model implementations, and in some cases provide extensions to make the models more generally applicable.     

Blood compatible N-maleyl chitosan-graft-PAMAM copolymer for enhanced gene transfection

To improve transfection efficiency, we prepared N-maleyl chitosan-graft-polyamidoamine (NMCTS-graft-PAMAM) copolymer. Self-assembled NMCTS-graft-PAMAM/pDNA complexes were prepared by complex coacervation method at different N/P (nitrogen to phosphate ratio) ratios. The copolymer effectively formed complexes with pDNA at lower N/P ratio (N/P ratio 1.0) than that of unmodified chitosan (N/P ratio 2.0) and the complexes were spherical with particle size of 100–150 nm. The copolymer showed significant protection of DNA from nuclease attack with lower toxicity against HeLa cell. The copolymer also showed no noticeable hemolytic effects up to 10 mg/mL indicating no detectable disturbance of the red blood cell membranes. The transfection efficiency of the copolymer was increased significantly compared to that of chitosan and reached up to 36 ± 2% at N/P ratio 7.0 which was higher than that of PEI (30 ± 3% at N/P ratio 10). Therefore, the copolymer may be a strong alternative candidate as effective nonviral vector.     

Versatile characterization of thiol-functionalized printed metal electrodes on flexible substrates for cheap diagnostic applications

Background

Cheap, reliable, point-of-care diagnostics is a necessity for the growing and aging population of the world. Paper substrate and printing method, combined together, are the cheapest possible method for generating high-volume diagnostic sensor platforms. Electrical transduction tools also minimize the cost and enhance the simplicity of the devices.

Methods

Standard surface characterization techniques, namely contact angle measurements, atomic force microscopy (AFM) and X-ray photoelectron spectroscopy (XPS) were used to analyze the growth of the organic thiol layers on top of the printed metal electrodes on paper substrates. The results were compared with those obtained by impedimetric electrical characterization method.

Results

This article reports the fabrication and characterization of printed metal electrodes and their functionalization by organic layers on paper and plastic substrates for biosensing and diagnostic applications. Impedimetric measurement is proposed as a simple, yet elegant, method of characterization of the organic layer growth.

Conclusions

Very good correlation was observed between the results of organic layer growth from different measurement methods, justifying the use of paper as a substrate, printing as a method for fabricating metal and organic layers and impedance as a suitable measurement method for hand-held diagnostic devices.

General significance

This result paves the way for the fabrication of more advanced bio-recognition layers for bio-affinity sensors using a printing technology that is compatible with flexible and cheap paper substrates. This article is part of a Special Issue entitled Organic Bioelectronics — Novel Applications in Biomedicine.     

Proton NMR Spectra of Peracetylated Derivatives of α,α-Trehalose and Sucrose in the Presence of a Europium Shift Reagent

The possibility of using recombinant human lactoferrin from rice (rhLF) makes it necessary to study its differences from the protein of milk. In this work, the binding of different iron-saturated forms of rhLF to Caco-2 cells was studied. Iron-saturated rhLF bound in higher proportion than the apo-form, but, the data obtained for specific binding were not compatible with receptor-mediated binding. Competition assays showed the same binding capacity for human milk lactoferrin as for rhLF to Caco-2 cells. Another basic protein of milk, lactoperoxidase, was found to compete with rhLF for binding to Caco-2 cell membranes, suggesting an electrostatic interaction. The transport of iron (⁵⁹Fe) bound to rhLF and to citrate and the transport of rhLF (¹²⁵I-labeled) were studied on Caco-2 monolayers. Transport of iron was found to be significantly greater when bound to citrate than to rhLF. The amount of intact lactoferrin that traversed the Caco-2 monolayers was very low, suggesting degradation of it across these cells.     

材料表面化学刺激对纤连蛋白构象及细胞的影响

生物材料的表面化学性质通过改变吸附蛋白的构象,影响细胞粘附铺展,产生不同的细胞行为.采用金-硫自组装单分子层技术(alkanethiol self-assembled monolayers,SAMs)构建了不同化学基团(-NH2、-COOH、-OH和-CH3)修饰的基底材料表面.运用X射线光能质谱(XPS)和接触角仪表...     

Force-field dependence on the liquid-expanded to liquid-condensed transition in DPPC monolayers

We have investigated using molecular dynamics simulations, the influence of the interaction cut-off and water model on the surface-pressure area isotherms of dipalmitoylphosphatidylcholine monolayers, where the phospholipids and the water molecules are modelled atomistically. We find that both the cut-off employed and the water model, influence the pressure area isotherms and the location of the liquid-expanded to liquid-condensed transitions. The combination of the Berger's et al. force field, with the TIP4P/2005 water model, and a long cut-off for the pair interactions ( ≥ 1.7 nm) provides a more accurate prediction of the surface pressure–area isotherm and reproduces the liquid condensed–liquid expanded transition observed in the experiments at 310 K.