Human olfactory receptor 17-40 as active part of a nanobiosensor: A microscopic investigation of its electrical properties

Increasing attention has been recently devoted to protein-based nanobiosensors. The main reason is the huge number of possible technological applications, going from drug detection to cancer early diagnosis. Their operating model is based on the protein activation and the corresponding conformational change, due to the capture of an external molecule, the so-called ligand. Recent measurements, performed with different techniques on human 17-40 olfactory receptor, evidenced a very narrow window of response in respect of the odour concentration. This is a crucial point for understanding whether the use of this olfactory receptor as sensitive part of a nanobiosensor is a good choice. In this paper we investigate the topological and electrical properties of the human olfactory receptor 17-40 with the objective of providing a microscopic interpretation of available experiments. To this purpose, we model the protein by means of a graph able to capture the mean features of the 3D backbone structure. The graph is then associated with an equivalent impedance network, able to evaluate the impedance spectra of the olfactory receptor, in its native and activated state. We assume a topological origin of the different protein electrical responses to different ligand concentrations: In this perspective all the experimental data are collected and interpreted satisfactorily within a unified scheme, also useful for application to other proteins.Comment: 6 pages, 6 figures, DOI:10.1039/c1ra0002

Experimental infection of Pacific oyster Crassostrea gigas spat by ostreid herpesvirus 1: demonstration of oyster spat susceptibility

In 2008 and 2009, acute mortalities occurred in France among Pacific cupped oyster, Crassostrea gigas, spat. Different hypothesis including the implication of environmental factors, toxic algae and/or pathogens have been explored. Diagnostic tests indicated that OsHV-1 including a particular genotype, termed OsHV-1 μVar, was detected in most of samples and especially in moribund oysters with the highlighting of virus particles looking like herpes viruses by TEM examination. In this study, an experimental protocol to reproduce OsHV-1 infection in laboratory conditions was developed. This protocol was based on the intramuscular injection of filtered (0.22 μm) tissue homogenates prepared from naturally OsHV-1 infected spat collected on French coasts during mortality outbreaks in 2008. Results of the experimental trials showed that mortalities were induced after injection. Moreover, filtered tissue homogenates induced mortalities whereas the same tissue homogenates exposed to an ultraviolet (UV) treatment did not induce any mortality suggesting that oyster spat mortalities require the presence of a UV sensitive agent. Furthermore, analysis of injected oyster spat revealed the detection of high amounts of OsHV-1 DNA by real-time quantitative PCR. Finally, TEM analysis demonstrated the presence of herpes virus particles. The developed protocol allowed to maintain sources of infective virus which can be useful for the development of further studies concerning the transmission and the development of OsHV-1 infection

Novel molecularly imprinted impedimetric biosensor based on polypyrrole and decorated graphene oxide for the routine monitoring of Lysozyme

In this work, a novel molecularly imprinted polymer based on polypyrrole (PPy) and decorated graphene oxide (GO@Fe3O4) was developed for the sensitive detection of lysozyme (LYS). The synthesized material (MIPPy/GO@Fe3O4) was electrodeposited with LYS as a template on gold microelectrodes. Transmission electron microscopy (TEM) and X-ray photoelectron spectroscopy (XPS) were used to confirm the adequate preparation of GO@Fe3O4, and the characterization of the resulting microsensors was carried out with the following analytical techniques: electrochemical impedance spectrometry (EIS), FT-IR analysis and scanning electron microscopy (SEM). An equivalent circuit was suggested to quantitatively analyse each component of the sensor system. EIS was also used for the determination of LYS in a wide linear range from 1 to 1 105 pg/mL, presenting good precision (RSD ≈ 10%, n = 5) and low limits of detection and quantification (LOD = 0.009 pg/mL and LOQ = 0.9 pg/mL, respectively). Meanwhile, the microsensor showed a high sensitivity, a good selectivity and reproducibility. The construction process was relatively simple, and provided a rapid and economical method for the routine monitoring of LYS. The microsensor was successfully applied for the detection of this protein in fresh chicken-egg white sample and commercial drug.Campus de Excelencia Internacional Andalucía Tech. Beca FPU18/05371 Proyecto de la Junta de Andalucía UMA18FEDERJA06

Los mundos [teóricos] de Coraline: Psicoanálisis, Postfeminismo y Postmodernismo en el cine de animación

[ES] Este artículo contribuye al estudio de diferentes metodologías como modo de aproximación al estudio teórico sobre la animación, utilizando como caso de estudio la película de animación Los Mundos de Coraline (Henry Selick, 2009), que incorpora a su complejo hilo argumental ideas procedentes de diversos movimientos histórico-sociales y filosóficos. Además, esta investigación justifica la necesidad de integrar en el actual cine de animación una fuerte inspiración teórica en la narrativa, además de las innovaciones tecnológicas que se desarrollan paralelamente. A través de la revisión de esta producción animada, se alega que los personajes animados pueden y deben ser protagonistas de narrativas tan profundas como las que aquí se tratan, ya que la animación tiene la potencialidad de incidir profundamente en cuestiones como la confirmación de la identidad o el desarrollo cognitivo del personaje protagonista, a través de la simbólica visualización de sus experiencias, miedos y frustraciones en el camino hacia su madurez.Martínez González, E. (2011). Los mundos [teóricos] de Coraline: Psicoanálisis, Postfeminismo y Postmodernismo en el cine de animación. Con A de Animación. (1):79-96. doi:10.4995/caa.2011.8627996

A Sensitive Micro Conductometric Ethanol Sensor Based on an Alcohol Dehydrogenase-Gold Nanoparticle Chitosan Composite

In this paper, a microconductometric sensor has been designed, based on a chitosan composite including alcohol dehydrogenase—and its cofactor—and gold nanoparticles, and was calibrated by differential measurements in the headspace of aqueous solutions of ethanol. The role of gold nanoparticles (GNPs) was crucial in improving the analytical performance of the ethanol sensor in terms of response time, sensitivity, selectivity, and reproducibility. The response time was reduced to 10 s, compared to 21 s without GNPs. The sensitivity was 416 µS/cm (v/v%)−1 which is 11.3 times higher than without GNPs. The selectivity factor versus methanol was 8.3, three times higher than without GNPs. The relative standard deviation (RSD) obtained with the same sensor was 2%, whereas it was found to be 12% without GNPs. When the air from the operator’s mouth was analyzed just after rinsing with an antiseptic mouthwash, the ethanol content was very high (3.5 v/v%). The background level was reached only after rinsing with water

Development of an ImmunoFET for Analysis of Tumour Necrosis Factor- (alfa) in Artificial Saliva: Application for Heart Failure Monitoring

Assessing tumour necrosis factor-(alfa) (TNF-(alfa)) levels in the human body has become an essential tool to recognize heart failure (HF). In this work, label-free, rapid, easy to use ImmunoFET based on an ion-sensitive field effect transistor (ISFET) was developed for the detection of TNF-(alfa) protein. Monoclonal anti-TNF-(alfa) antibodies (anti-TNF-(alfa) mAb) were immobilized on an ISFET gate made of silicon nitride (Si3N4) after salinization with 11-(triethoxysilyl) undecanal (TESUD). The obtained ISFET functionalized with the mAbs (ImmunoFET) was used to detect TNF-(alfa) protein in both phosphate buffer saline (PBS) and artificial saliva (AS). The change in the threshold voltage of the gate (DVT) showed approximately linear dependency on the concentration of the antigens in the range 5-20 pg/mL for both matrixes. The cross-selectivity study showed that the developed ImmunoFET demonstrated to be selective towards TNF-(alfa), when compared to other HF biomarkers such as N-terminal pro-brain natriuretic peptide (NT-proBNP), interleukin-10 (IL-10), and cortisol, even if further experiments have to be carried out for decreasing possible unspecific absorption phenomena. To the best of our knowledge, this is the first ImmunoFET that has been developed based on Si3N4 for TNF-(alfa) detection in AS by electrical measurement

Electrochemical Boron-Doped Diamond Film Microcells Micromachined with Femtosecond Laser: Application to the Determination of Water Framework Directive Metals

Planar electrochemical microcells were micromachined in a microcrystalline boron-doped diamond (BDD) thin layer using a femtosecond laser (Photo 1). The electrochemical performances of the new laser-machined BDD microcell were assessed by differential pulse anodic stripping voltammetry (DPASV) determinations, at nM level, of the four heavy metal ions of the European Water Framework Directive (WFD): Cd(II), Ni(II), Pb(II), Hg(II). The results are compared with those of previously published BDD electrodes [1]. The calculated detection limits are 0.4 nM, 6.8 nM and 5.5 nm 2.3 nM, and the linearities go up to 35nM, 97nM, 48nM and 5nM for respectively Cd(II), Ni(II) Pb(II) and Hg(II). The detection limits meet with the environmental quality standard of the WFD for three of the four metals. It was shown that the four heavy metals could be detected simultaneously, in the concentration ratio usually measured in sewage or runoff waters

Recent Trends in Monitoring of European Water Framework Directive Priority Substances Using Micro-Sensors: A 2007–2009 Review

This review discusses from a critical perspective the development of new sensors for the measurement of priority pollutants targeted in the E.U. Water Framework Directive. Significant advances are reported in the paper and their advantages and limitations are also discussed. Future perspectives in this area are also pointed out in the conclusions. This review covers publications appeared since December 2006 (the publication date of the Swift report). Among priority substances, sensors for monitoring the four WFD metals represent 81% of published papers. None of analyzed publications present a micro-sensor totally validated in laboratory, ready for tests under real conditions in the field. The researches are mainly focused on the sensing part of the micro-sensors. Nevertheless, the main factor limiting micro-sensor applications in the environment is the ruggedness of the receptor towards environmental conditions. This point constitutes the first technological obstacle to be overcome for any long-term field tests