Nanobodies and Nanocrystals: Highly Sensitive Quantum Dot‐Based Homogeneous FRET Immunoassay for Serum‐Based EGFR Detection

Semiconductor quantum dot nanocrystals (QDs) for optical biosensing applications often contain thick polyethylene glycol (PEG)‐based coatings in order to retain the advantageous QD properties in biological media such as blood, serum or plasma. On the other hand, the application of QDs in Förster resonance energy transfer (FRET) immunoassays, one of the most sensitive and most common fluorescence‐based techniques for non‐competitive homogeneous biomarker diagnostics, is limited by such thick coatings due to the increased donor‐acceptor distance. In particular, the combination with large IgG antibodies usually leads to distances well beyond the common FRET range of approximately 1 to 10 nm. Herein, time‐gated detection of Tb‐to‐QD FRET for background suppression and an increased FRET range is combined with single domain antibodies (or nanobodies) for a reduced distance in order to realize highly sensitive QD‐based FRET immunoassays. The “(nano)²” immunoassay (combination of nanocrystals and nanobodies) is performed on a commercial clinical fluorescence plate reader and provides sub‐nanomolar (few ng/mL) detection limits of soluble epidermal growth factor receptor (EGFR) in 50 μL buffer or serum samples. Apart from the first demonstration of using nanobodies for FRET‐based immunoassays, the extremely low and clinically relevant detection limits of EGFR demonstrate the direct applicability of the (nano)^2‐ assay to fast and sensitive biomarker detection in clinical diagnostics.     

Structure, infrared and Raman spectroscopic studies of new Sr0.50SbFe(PO4)3 and SrSb0.50Fe1.50(PO4)3 Nasicon phases

Two newly synthesised Sr_0.50SbFe(PO₄)₃ [Sr_0.5.] and SrSb_0.50Fe_1.50(PO₄)₃ [Sr.] phases were obtained by conventional solid-state reaction techniques at 1000 °C in air atmosphere. Their crystallographic structures were determined at room temperature from X-ray powder diffraction (XRPD) data using the Rietveld analysis. Both compounds belong to the Nasicon structural family. [Sr_0.5.] and [Sr.] crystallise in rhombohedral system with textR[`3] {text{R}}overline{3} and textR[`3] textc {text{R}}overline{3} {text{c}} space group, respectively. Hexagonal cell parameters for [Sr_0.5.] and [Sr.] are: a = 8.227(1) ?, c = 22.767(2) ? and a = 8.339(1) ?, c = 22.704(2) ?, respectively. Sr²⁺ and vacancies in {[Sr_0.50]_3a[□_0.50]_3b}_M1SbFe(PO₄)₃ are practically ordered within the two positions, 3a and 3b, of M1 sites. Structure refinements show also an ordered distribution of Sb⁵⁺ and Fe³⁺ ions within the Nasicon framework. Within the structure, each Sr_(3a)O₆ octahedron shares two faces with two Fe³⁺O₆ octahedra and each vacancy (□_(3b)O₆) site is located between two Sb⁵⁺O₆ octahedra. In [Sr]_M1Sb_0.50Fe_1.50(PO₄)₃ compound, all M1 sites are occupied by Sr²⁺ and the Sb⁵⁺ and Fe³⁺ ions are randomly distributed within the Nasicon framework. A Raman and infrared spectroscopic study was used to obtain further structural information about the nature of bonding in both selected compositions.     

Co0.5TiOPO4: Crystal structure, magnetic and electrochemical properties

The new titanium oxyphosphate Co_0.5TiPO₅ has been prepared by solid state reaction. Its structure has been determined by single crystal X-ray diffraction and was further investigated by FT-IR spectroscopy and magnetic measurements. The compound crystallizes in the monoclinic system, S.G: P2₁/c [a = 7.358(1) Å, b = 7.378(2) Å, c = 7.383(3) Å, β = 119.66(3)°, Z = 4, R₁ = 0.0142, wR₂ = 0.0429]. The structure can be described as a network of very distorted TiO₆ octahedra, in which the Ti⁴⁺ ions are displaced from the centres of the octahedra, and slightly distorted PO₄ tetrahedra. Half of the octahedral cavities are occupied by Co atoms. The other half of octahedral sites is vacant and favourable for the electrochemical insertion of lithium. The insertion of lithium was studied by galvanostatic charging and discharging between different voltage limits.     

Measured infrared optical cross sections for a variety of chemical and biological aerosol simulants

We conducted a series of spectral extinction measurements on a variety of aerosolized chemical and biological simulants over the spectral range 3-13 microm using conventional Fourier-transform IR (FTIR) aerosol spectroscopy. Samples consist of both aerosolized particulates and atomized liquids. Materials considered include Bacillus subtilis endospores, lyophilized ovalbumin, polyethylene glycol, dimethicone (SF-96), and three common background materials: kaolin clay (hydrated aluminum silicate), Arizona road dust (primarily SiO2), and diesel soot. Aerosol size distributions and mass density were measured simultaneously with the FTIR spectra. As a result, all optical parameters presented here are mass normalized, i.e., in square meters per gram. In an effort to establish the utility of using Mie theory to predict such parameters, we conducted a series of calculations. For materials in which the complex indices of refraction are known, e.g., silicone oil (SF-96) and kaolin, measured size distributions were convolved with Mie theory and the resultant spectral extinction calculated. Where there was good agreement between measured and calculated extinction spectra, absorption, total scattering, and backscatter were also calculated.     

Anisotropy of hygrothermal damage in fiber/polymer composites: Effective elasticity measures and estimates

Measured elasticity moduli of a highly (68%) glass-fiber reinforced epoxy matrix for different amounts of fiber/matrix interface weakening and debonding, due to different hygrothermal ageing stages, are compared to estimated ones. Ultrasonic measurements provide seven of the nine elasticity moduli of the orthotropic material samples, including all the moduli significantly affected by damage. Theoretical estimates combine homogenization modeling techniques and Finite Element (FE) calculations, the latter when the effect of observed partial debonding on effective moduli is to be specified. These estimates are performed under different assumptions for the composite structure, with special attention to the existence of a fiber–matrix interphase. Analytical comparisons for the undamaged composite establish that matching US measurements with estimates cannot be obtained, regardless of the chosen model, without the assumption of an interphase layer of modified resin coating the fibers. This coating resin, when in relevant concentration with regard to literature data about fiber coating thickness, typically conserves the epoxy moduli transversally to the fiber orientation, while, in the fiber direction its moduli approach those of the fibers. The comparison of the US measurements on damaged samples to FE calculations assuming progressive one-directional debonding shows that most of the composite stiffness loss can preferentially be due to an initial interphase weakening, while the fiber/matrix debonding seems more likely delayed to long H-ageing times. This is consistent with physical interpretation of damage by water pooling through silane bridges bonding epoxy to glass. The calculations also provide the effective stiffness, at different damage stages, of the “Undamaged Equivalent Inhomogeneity” for this damaged inclusion type.     

Definition of the diffusion coefficient in scattering and absorbing media

We revisit the definition of the diffusion coefficient for light transport in scattering and absorbing media. From an asymptotic analysis of the transport equation, we present a novel derivation of the diffusion coefficient, which is restricted neither to low absorption nor to a situation in which the specific intensity is quasi-isotropic. Our result agrees with previous expressions of the diffusion coefficient in the appropriate limit. Using numerical simulations, we discuss the implications of the proper choice of the diffusion coefficient for time-dependent transport.     

Evaluation of a program to reduce motor-vehicle collisions among young adults in the county of Landes, France

The aim of this study was to evaluate the impact of a prevention program called “Atout-Route”, based on the concept of commitment. The program was implemented in March 2000, to reduce the number of drivers under the age of 25 years involved in motor-vehicle collisions in the county of Landes (southwestern France). Using data from the regional observatory of road safety, we defined a target and three control groups similar on age or location. We used Poisson and quasi-Poisson regression to estimate whether the observed evolution of motor-vehicle collisions, after the program was implemented, was different in the targeted group than in the three control groups. The number of motor-vehicle collisions decreased everywhere and in every age group. The effect of the prevention program was not statistically significant (relative risk = 0.89; 95% confidence interval 0.74–1.07). Our results are compatible with a positive effect of the program. The possible dilution of its effect by national road safety actions implemented since 2000, and our early assessment are possible reasons for the non-significant observation.