Impact of Collection Volume and DNA Extraction Method on the Detection of Biomarkers and HPV DNA in First-Void Urine

The potential of first-void (FV) urine as a non-invasive liquid biopsy for detection of human papillomavirus (HPV) DNA and other biomarkers has been increasingly recognized over the past decade. In this study, we investigated whether the volume of this initial urine stream has an impact on the analytical performance of biomarkers. In parallel, we evaluated different DNA extraction protocols and introduced an internal control in the urine preservative. Twenty-five women, diagnosed with high-risk HPV, provided three home-collected FV urine samples using three FV urine collection devices (Colli-Pee) with collector tubes that differ in volume (4, 10, 20 mL). Each collector tube was prefilled with Urine Conservation Medium spiked with phocine herpesvirus 1 (PhHV-1) DNA as internal control. Five different DNA extraction protocols were compared, followed by PCR for GAPDH and PhHV-1 (qPCR), HPV DNA, and HBB (HPV-Risk Assay), and ACTB (methylation-specific qPCR). Results showed limited effects of collection volume on human and HPV DNA endpoints. In contrast, significant variations in yield for human endpoints were observed for different DNA extraction methods (p < 0.05). Additionally, the potential of PhHV-1 as internal control to monitor FV urine collection, storage, and processing was demonstrated.     

Quantification of Tin and Lead in Binary Alloys Using Voltammetry of Immobilized Microparticles

Voltammetry of immobilized microparticles (VMP) has been used in this work for the quantitative determination of tin and lead particles in their binary alloys. Carbon paste electrodes, which contained small amounts of tin and lead or their mixtures, were used as working electrodes and square wave voltammograms of each electrode were recorded. Quantification was performed using optimum experimental conditions, obtained by an experimental design technique. The calibration was made by measuring the percentage peak height of each metal taking the sum of peak currents of the both metals as 100 %. The results were compared with quantitative results of X‐ray fluorescence (XRF) technique to evaluate the capability of VMP method in its quantitative determination of solid samples.     

Microcapillary electrochemical droplet cells: applications in solid-state surface analysis

Capillary–based microcells, also known as microcapillary electrochemical droplet cells, have proved their capabilities in various electrochemical surface investigations in recent decades. Due to the large measured current density and the high limiting current, this technique provides high–resolution electrochemical responses. Current densities in the range from a few femto to pico Acm^?2 to hundreds of Acm^?2 can be measured using this technique. Various applications for microcapillary cells have been reported. Technical limitations, such as the Ohmic drop and changes in the composition of the measurement area near the tip of the microcapillary have also been considered by some researchers. The rapid increase in the application of microcells and the increase in the number of related reports published in the literature have paralleled recent attempts to develop and improve microcell setups, showing that this technique is already well established for electrochemical surface studies.     

Electrochemistry reveals archaeological materials

The characterization of materials constituting cultural artefacts is a challenging step in their conservation, due to the object’s uniqueness and the reduced number of conservation institutes able to supply non-destructive analysis. We propose an alternative analytical tool, which combines accessibility (low cost and portable) and high sensitivity, based on electrochemical linear sweep voltammetry (LSV) with paraffin impregnated graphite electrode (PIGE). To investigate the composition of “white alloys” that certainly have been used as decoration on copper-based Roman fibulae, sampling was done very locally by gently rubbing the selected areas with the PIGE. LSV results evidence the presence of silver, lead, and tin, supporting the argument provided by typological analysis that these metals were used for decoration.     

Surface characterization of a cross‐linked cytochrome c film on cysteamine‐modified gold electrodes

The aim of the present paper is to characterize a cross‐linked horse heart cytochrome c (HHC) film on cysteamine‐modified gold electrodes. The HHC film was deposited using 1‐ethyl‐3‐(3‐dimethylaminopropyl)‐carbodiimide (EDC) as a coupling agent. Attenuated total reflection infrared (ATR‐IR) spectroscopic analyses were performed to characterize the newly formed surface on a qualitative and conformational level. The film thickness was measured using a noncontact optical surface profiler, while quantitative data and information on the heterogeneity of the film were obtained by means of synchrotron radiation X‐ray micro fluorescence (SR micro‐XRF). Results indicate that, in addition to electrochemical studies, spectroscopic analysis methods are essential to gain insight in the effect of immobilization strategies on protein conformations. The latter is of relevance in the development and optimization of biosensors. Copyright © 2009 John Wiley & Sons, Ltd.     

Oxidation-sensitive polymeric nanoparticles

We have recently demonstrated the possible use of organic polysulfides for the design of oxidation-sensitive colloidal carriers in the form of polymeric vesicles, which are particularly suitable for the encapsulation of hydrosoluble drugs. In the present research we extend our efforts to carriers specifically suitable for hydrophobic molecules. Exploiting the living emulsion polymerization of episulfides, we have produced new cross-linked polysulfide nanoparticles. Here we demonstrate how this process allows the production of stable nanoparticles with a good control over their size and functionality. The nanoparticles showed negligible cytotoxicity on a fibroblast model; furthermore, they exhibited sensitivity to oxidative conditions, which first produce swelling and then solubilize the material.     

Strategy for the synthesis of multivalent peptide-based nonsymmetric dendrimers by native chemical ligation

A strategy for the synthesis of multivalent peptide-based nonsymmetric dendrimers by native chemical ligation using poly(lysine) dendritic wedges as scaffolds is presented.     

Double pulse laser ablation and laser induced breakdown spectroscopy: A modeling investigation

A numerical model, describing laser–solid interaction (i.e., metal target heating, melting and vaporization), vapor plume expansion, plasma formation and laser–plasma interaction, is applied to describe the effects of double pulse (DP) laser ablation and laser induced breakdown spectroscopy (LIBS). Because the model is limited to plume expansion times in the order of (a few) 100 ns in order to produce realistic results, the interpulse delay times are varied between 10 and 100 ns, and the results are compared to the behavior of a single pulse (SP) with the same total energy. It is found that the surface temperature at the maximum is a bit lower in the DP configuration, because of the lower irradiance of one laser pulse, but it remains high during a longer time, because it rises again upon the second laser pulse. Consequently, the target remains for a longer time in the molten state, which suggests that laser ablation in the DP configuration might be more efficient, through the mechanism of splashing of the molten target. The total laser absorption in the plasma is also calculated to be clearly lower in the DP configuration, so that more laser energy can reach the target and give rise to laser ablation. Finally, it is observed that the plume expansion dynamics is characterized by two separate waves, the first one originating from the first laser pulse, and the second (higher) one as a result of the second laser pulse. Initially, the plasma temperature and electron density are somewhat lower than in the SP case, due to the lower energy of one laser pulse. However, they rise again upon the second laser pulse, and after 200 ns, they are therefore somewhat higher than in the SP case. This is especially true for the longer interpulse delay times, and it is expected that these trends will be continued for longer delay times in the μs-range, which are most typically used in DP LIBS, resulting in more intense emission intensities.