Mechanoelectrochemical synthesis of porous Ti-based nanocomposite biomaterials

In this work we have synthesized a new class of nanocomposites based on Ti with the addition of hydroxyapatite (HA) and glass 45S5. The nanocomposites were prepared by mechanical alloying of the pure microcrystalline Ti powders with different amount of ceramics. The powder mixture was milled up to 48 h, pressed and sintered, which resulted in nanocomposite structure with the grain size of about 20–36 nm. The ultra low grain size structure improves mechanical properties of the implants in comparison to commonly used microcrystalline Ti-based implants. For example, the hardness of the Ti-HA nanocomposites reaches a value of 1500 HV and is five times greater than the microcrystalline Ti.To improve bonding of the implants with human tissue, the implants were electrochemically etched in 1 M H₃PO₄ + 2–10% HF electrolyte at 10 V vs. OCP for times up to 60 min. The treatment results in highly porous surface covered with Ti-oxide. The nanocrystalline structure is very useful during etching, due to the easy access of the electrolyte to the large volume of the grain boundaries. The nanocomposites with modified surface show very good corrosion resistance in Ringer’s solution.     

Perturbation of Conformational Dynamics of ASCUT-1 from Ascaris lumbricoides by Temperature and Sodium Dodecyl Sulfate

ASCUT-1 is a protein found in cuticlin, the insoluble residue of the cuticles of the nematode Ascaris lumbricoides. It contains the CUT-1-like domain which is shared by members of a novel family of components of extracellular matrices. The monomeric form of ASCUT-1 contains a single tryptophan residue. An understanding of the structure-function relationship of the protein under different chemical-physical conditions is of fundamental importance for an understanding of its structure and function in cuticles. In this paper we report the effect of the temperature and sodium dodecyl sulfate on the structural stability of this protein. The structure of the protein was studied in the temperature range 25–85°C in the absence and in the presence of sodium dodecyl sulfate by frequency-domain measurements of the intrinsic fluorescence intensity and anisotropy decays. The time-resolved fluorescence data in the absence of SDS indicated that the tryptophanyl emission decays were well described by a bimodal lifetime distribution, and that the temperature increases resulted in the sharpening and in the shortening of the tryptophanyl lifetime distribution. In the presence of SDS an unimodal fluorescence lifetime distribution as well as a marked decrease in the anisotropy decay values were observed.     

Adsorption kinetics of p-nitrobenzenethiol self-assembled monolayers on a gold surface

The adsorption kinetics of octadecanethiol (ODT) and p-nitrobenzenethiol (NBT) from ethanol solutions has been studied by means of contact angle, optical ellipsometry, angle-resolved X-ray photoelectron spectroscopy (ARXPS), and grazing angle attenuated total reflection Fourier transform infrared (FTIR) measurements. ODT data were used as a reference for the kinetics studies of film growth. The growth of self-assembled monolayers from dilute solutions follows Langmuir isotherm adsorption kinetics. A saturated film is formed within 5 h after immersion in solutions of concentrations ranging from 0.0005 to 0.01 mM. The density of the monolayer depends on the concentration of the solution.     

Förster energy transfer from nonexponentially decaying donors

Necessary modifications to the expression for the F?rster energy transfer rate are discussed when fluorescence decay of the donor in the absence of acceptor is nonexponential. Discrete and continuous models of the nonexponentiality are taken into account. No general solution of the problem is found. It is, however, suggested that in many of the biochemical problems the most appropriate modification of the transfer rate can be that which is based on the assumption of the same constant value of the radiative decay rate for all donor molecules. The effect of the assumed form of the F?rster energy transfer rate on the recovered values of the distance distribution and dynamics parameters of some exemplary bichromophoric systems is examined.     

Exploring fundamental differences between red- and blue-shifted intramolecular hydrogen bonds using FAMSEC,FALDI, IQA and QTAIM

We have discovered, using developed by us recently FALDI and FAMSEC computational techniques, fundamentally distinct mechanisms of intramolecular red- and blue-shifted H-bond formation that occurred in different conformers of the same molecule (amino-acid β-alanine) involving the same heteroatoms (O–H???N and N–H???O). Quantitative topological, geometric and energetic data of both H-bonds obtained with well-known QTAIM and IQA methodologies agree with what is known regarding H-bonding in general. However, the FALDI charge and decomposition scheme for calculating in real space 3D conformational deformation densities provided clear evidence that the process of electron density redistribution taking place on the formation of the stronger red-shifted H-bond is fundamentally distinct from the weaker blue-shifted H-bond. Contributions made by atoms of the X–H???Y–Z fragment (IUPAC notation) as well as distinct atoms on the H-bond formation were fully explored. The FAMSEC energy decomposition approach showed that the atoms involved in formation of the red-shifted H-bond interact in a fundamentally different fashion, both locally and with the remainder of the molecule, as compared with those of the blue-shifted H-bond. Excellent correlations of trends obtained with QTAIM, IQA, FAMSEC and FALDI techniques were obtained. Commentary regarding IUPAC recommended definition of an H-bond and validity of observed AILs (or bond paths) of the two H-bond kinds is also discussed.     

Silver nanoparticle-enhanced fluorescence in microtransponder-based immuno- and DNAhybridization assays

The aim of this study is to improve assay sensitivity in common solid-phase bioassay configurations as the result of using silver nanoparticles. The solid phase was provided by numerically indexed, silicon-based electronic chips, microtransponders (p-Chips) that have previously been used in multiplexed assays. Assay configurations investigated included an ELISA-type immunoassay and a DNA hybridization assay. The surface of p-Chips was derivatized with the silver island film (SIF) and a polymer, and then characterized with AFM and SEM. Silver nanoparticle sizes were in the range of 100 to 200 nm. Four fluorophores were tested for fluorescence enhancement; namely, green fluorescent protein, phycoerythrin, Cy3 and Alexa Fluor 555. We consistently observed significant fluorescence enhancement and sensitivity improvement in the p-Chip-based assays: the sensitivity in the cytokine IL-6 immunoassay was 4.3 pg/ml, which represented a 25-fold increase over the method not involving a SIF; and 50 pM in the hybridization assay, a 38-fold increase. The greatest enhancement was obtained for p-Chip surfaces derivatized first with the polymer and then coated with SIF. In conclusion, we show that the SIF-p-Chip-based platform is a highly sensitive method to quantify low-abundance biomolecules in nucleic acid-based assays and immunoassays.     

Surface plasmon-coupled polarized emission of N-acetyl-l-tryptophanamide

We report an observation of ultraviolet (UV) surface plasmon-coupled emission (SPCE) of N-acetyl-l-tryptophanamide (NATA). The sample was spin coated from poly(vinyl alcohol) (PVA) solution on 20 nm aluminum film deposited on a quartz substrate. The directional UV SPCE occurs within a well-defined narrow angle at 52 degrees from the normal to the coupling hemicylinder quartz prism. The NATA directional emission is highly p polarized as expected for surface plasmon-coupled radiation. The 10 nm protective SiO2 layer deposited on top of the aluminum film significantly neutralized the fluorophore quenching by the metal surface. SPCE of NATA demonstrates a remarkable intrinsic dispersive property-the maximum of the emission spectrum depends on the observation angle. The efficient spectral resolution of SPCE can be used in the construction of miniaturized spectrofluorometers. The observation of SPCE of tryptophan opens a new possibility for the study of many unlabeled proteins with the technique complementary to surface plasmon resonance analysis.     

Voltammetry as Virtual Potentiometric Sensor in Modelling of a Metal/Ligand System and Refinement of Stability Constants. Part 5

The concept of virtual potential (employed here in modelling operations), a unique experimental setup designed and built in our laboratories, and new regression equations derived for nonlinear fitting of quasi‐reversible direct‐current polarograms were combined with the existing rigorous treatment and refinement of polarographic data to establish reliable metal/ligand models and accurate stability constants for the lead(II)/glycine/OH^? and lead(II)/sarcosine/OH^? systems (sarcosine = N‐methylglycine). In the case of glycine, the complexes [M(HL)], [ML], [ML₂], and [ML₃] were identified, and their stability constants (as log β) were established to be 10.51 ± 0.06, 4.58 ± 0.02, 7.19 ± 0.10, and 9.27 ± 0.02, respectively, the complex [ML₃] being reported here for the first time (Table 2). The system with sarcosine involving [M(HL)], [ML], [ML₂], [ML₃], and [ML₂(OH)₂], with the stability constants (as log β) 11.01 ± 0.04, 4.18 ± 0.03, 7.23 ± 0.03, 9.1 ± 0.3, and 15.97 ± 0.07, respectively, is reported for the first time (Table 3). The log K₁ value for Pb^II with sarcosine is a fraction of a log unit smaller when compared with the Pb^II complex with glycine, in agreement with the literature data for Cu^II, Ni^II, and Zn^II showing the same trend for these two ligands. The proposed nonlinear curve‐fitting operations expand the applicability of polarography to study reliably and conveniently quasi‐reversible, on the polarographic time scale, metal/ligand systems (systems with involved heterogeneous kinetics).