Analyse des mutations des domaines ISDR et V3 de la protéine NS5A du virus de l'hépatite C avant le traitement par l'interféron avec ou sans ribavirine

Aim of the study. – The hepatitis C virus (HCV) non-structural NS5A protein has been controversially implicated in the resistance of HCV to interferon therapy in clinical studies. In Japan, mutations in the interferon sensitivity-determining region (ISDR) in the NS5A gene were associated with response to interferon therapy in patients infected with genotype 1b. In contrast, studies from Europe did not confirm such association. More recently, it has been suggested that the V3 domain outside the putative ISDR might also have amino acids changes that may be involved in the resistance to IFN. In this study, the relationship between NS5A mutations in ISDR and V3 domains and virological response to therapy were investigated. Materials and methods. – The NS5A gene was sequenced from 35 HCV genotype 1b infected patients at D0 of a prospective clinical trial of interferon therapy and interferon plus Ribavirin combination therapy. Results. – In the ISDR domain, we did not observe any significant differences in amino acids changes between responders (1.7 ± 1.8, n = 19, range 0–6) and non-responders (1.1 ± 0.8, n = 14, range: 0–3), (P = 0.483), to therapy before the beginning of treatment. In the V3 domain, we found more mutations in responders (6.5 ± 1.9, range: 2–11) than in non-responders (4.7 ± 1.2, range: 3–8) (P = 0.0013), before the beginning of treatment. Conclusion. – Our results confirm that, in Europe, the ISDR domain is not predictive for treatment success but suggest that the V3 domain have greater variability in responders than non-responders

Mutations within the hepatitis C virus genotype 1b E2-PePHD domain do not correlate with treatment outcome.

The hepatitis C virus (HCV) envelope protein 2 (E2) interacts in vitro with the interferon alpha (IFN-alpha)-inducible double-stranded RNA-activated protein kinase, suggesting a possible mechanism by which HCV may evade the antiviral effects of IFN-alpha. Variability in the part of the HCV E2 gene encoding the carboxy-terminal part of the protein, which includes the interaction domain (E2-PePHD), was explored in 25 patients infected with HCV genotype 1b and receiving IFN-alpha therapy. PCR products were generated and sequenced for 15 patients with a sustained response and for 10 patients with no virological response after treatment with IFN-alpha and ribavirin. PePHD amino acid sequences were obtained for isolates from serum collected before and during treatment, after 2 months in responders, and after 6 months in nonresponders. Quasispecies analysis of the pretreatment PePHD region was performed for isolates from patients displaying amino acid substitutions in this domain on direct sequencing. The E2-PePHD sequence was highly conserved in both resistant and susceptible genotype 1b strains and was identical to the prototype HCV type J sequence. No significant emergence of PePHD mutants during therapy was observed in our clonal analysis, and sporadic mutations and treatment outcomes were not found to be correlated. The PePHD sequence before or during treatment cannot be used to predict reliably the outcome of treatment in HCV type 1b-infected patients

Atomic environment and interfacial structural order of TiAlN/Mo multilayers

Multilayered TiAlN/Mo coatings were deposited by dc reactive magnetron sputtering in a custom-made chamber. In order to assess the composition of these coatings, a combined study of Extended X-ray Absorption Fine Structure (EXAFS) and Rutherford Backscattering Spectrometry (RBS) experiments were performed. Through the simulation of the EXAFS spectra, giving the local environment of the titanium atoms inside the nitride (TiAlN), a cubic phase has been evidenced with aluminium atoms occupying titanium sites. For modulation periods in the range of 3.6–11.8 nm, RBS simulations on these multilayers also enabled the determination of the level of intermixing that occurs at the interfaces as a function of the negative bias voltage and number of layers. It was observed that the intermixing width could be as high as 2.1 nm for the roughest samples (larger periods) or as low as 0.4 nm for those with the sharpest interfaces (smaller periods).Comunidade Europeia (CE). Fundo Europeu de Desenvolvimento Regional (FEDER) -Ministério da Ciência e Tecnologia.Cooperação Científica e Tecnológica Internacional (ICCTI) - Ambassade de France in Portugal - Project no. 543 B3/2001.Fundação para a Ciência e a Tecnologia (FCT) - Programa Operacional “Ciência, Tecnologia, Inovação - POCTI/32670/CTM/2000

X-Ray synchronotron study of phase transforms in illite clays to extract information on sigillata manufacturing processes.

The technique of sigillata really began in central Italy during the first century B. C. with the development of red vitrified slips obtained through vitrification of a clay preparation. These ceramics, usually decorated with raised motifs and standardised shapes, quickly took over as semi luxury crockery. Given this success, this technique quickly extended to the entire Italian peninsula and then to the Mediterranean coast. From the very start of our era, great centres of production were set up in the south of Gaul. The aspect of sigillata comes from the nature and the texture of its slip. Studies have shown that sigillata slips of quality were obtained from a non calcareous clay while the local calcareous clay was used for the bodies. During firing the slips are vitrified and get a specific microstructure containing hematite and nanometric corundum crystals [1]. An investigation of the clays surrounding La Graufesenque site started and it seems that only the Trias levels are chemically compatible with the composition of antique slips. Apart from the in depth study of the mineralogical nature of these clays realized at a geological Laboratory, we have studied the structural transformations as a function of temperature of two of these clays, chosen for the quality of vitrification in the firing temperature range of sigillata [1030-1080°C]. The main difference between the chemical composition of these two clays is the amount of Mg (2.4 % and 4.5 % in oxide weight). Time-resolved measurements were made at Daresbury (station 2.3) up to 1100oC in oxidizing conditions. An abrupt increase of the hematite cell was observed around 850°C. Above 1000°C, the hematite peaks get sharper which indicate an increase of coherence length (Fig. 1). A spinel phase with cell parameter close to MgAl2O3 was detected from this temperature. As for the hematite, its coherence length increases with the temperature but also during the beginning of the cooling. For the clay sample with the smaller amount of Mg, a corundum phase with very small coherence length was detected above 1000°C. Slips were prepared from the last clay by modern potters and firing at 1050°C in oxidizing atmosphere. A mineral quantitative analysis performed using the Rietveld method revealed that the amount of spinel phase is very high while the corundum contributes to a small part of crystal phases. It is the inverse in the antique slip where the amount of Mg in oxide weight is around 1%. It is clear that the amount of Mg plays a key role in the corundum/spinel competition and that the present slips contain too much Mg. Two questions arise: (i) As the Trias levels are quite heterogeneous is it possible to find clay with less Mg? and (ii) Did the gallo-roman potters eliminated a great part of Mg during the slip preparation process? We discuss the merits of these two alternative hypotheses

Structural evolution in ZrNxOy thin films as a function of temperature

Single-layered zirconium oxynitride (ZrNxOy) thin films have been deposited on steel substrates, at a constant temperature of 300 °C, by radiofrequency (rf) reactive magnetron sputtering of a pure Zr target in an argon-oxygen-nitrogen atmosphere. The variation of the flow rate of the reactive gases enabled changes in the composition and structure of the films. X-ray diffraction (XRD) and glancing incidence X-ray diffraction (GIXRD) were used to study the as-deposited films and their structural changes during or after heat treatment, from 400 to 900 °C, in controlled atmosphere and in vacuum.http://www.sciencedirect.com/science/article/B6TVV-4DPYN97-6/1/3785b40b130ad12af7221c230d2968c

Structural analysis of Ti1−xSixNy nanocomposite films prepared by reactive magnetron sputtering

Nanocomposite thin films consisting of both nanosized solid solutions or nanosized polycrystalline materials embedded in various amorphous matrix materials thus provide a grate potential for future mechanical devices. In this paper we report on the preparation of films resulting from additions of Si to TiN matrix by r.f. reactive magnetron sputtering. Structural properties such as growing characteristics (type of matrix, texture and grain size) will be analysed in some detail. Conventional transmission electron microscopy (TEM) and High-resolution transmission electron microscopy (HRTEM), together with both symmetric and asymmetric mode X-ray diffraction (XRD) experiments were used for this characterisation. The atomic composition of the samples was obtained by Rutherford Backscattering Spectrometry (RBS). The analysis will be carried out as a function of the Si content in the Ti1-xSixNy matrix and several relations will be made regarding important parameters such as texture evolution, grain sizes, but most specially by the type of matrix developed. Regarding the results, all samples develop a double fcc phase with lattice parameters of 4.30 Å and 4.17 Å, corresponding to cubic TiN and most likely to a cubic lattice of SiNx, respectively. Although no significant changes in texture were observed till Si compositions up to 10.6 at. %, the arrangement in atoms planes seem to vary. Results show that an asymmetric arrangement is developed in samples of small Si additions. This arrangement becomes more isotropic with the increase of Si contents, which we attribute to the increase in the SiNx content. In fact this increase leads also to the development of a small amorphous tissue of Si3N4 for large Si additions.The authors gratefully acknowledge the financial support of the ‘Fundação para a Ciência e Tecnologia’ (FCT) during the course of this research under project n PBICT/P/CTM/1962/95 and also the French CNRS Institution and the Portugese ICCTI Institution through CNRS/ICCTI Programs (N: 5522-1998 and 7087-1999)

Bimodal Electrochemiluminescence Microscopy of Single Cells

Electrochemiluminescence (ECL) microscopy is an emerging technique with new applications such as imaging of single entities and cells. Herein, we have developed a bimodal and bicolor approach to record both positive ECL (PECL: light-emitting object on dark background) and shadow label-free ECL (SECL: nonemissive object shadowing the background luminescence) images of single cells. This bimodal approach is the result of the simultaneous emissions of [Ru(bpy)3]2+ used to label the cellular membrane (PECL) and [Ir(sppy)3]3- dissolved in solution (SECL). By spectrally resolving the ECL emission wavelengths, we recorded the images of the same cells in both PECL and SECL modes using the [Ru(bpy)3]2+ (?max = 620 nm) and [Ir(sppy)3]3- (?max = 515 nm) luminescence, respectively. PECL shows the distribution of the [Ru(bpy)3]2+ labels attached to the cellular membrane, whereas SECL reflects the local diffusional hindrance of the ECL reagents by each cell. The high sensitivity and surface-confined features of the reported approach are demonstrated by imaging cell-cell contacts during the mitosis process. Furthermore, the comparison of PECL and SECL images demonstrates the differential diffusion of tri-n-propylamine and [Ir(sppy)3]3- through the permeabilized cell membranes. Consequently, this dual approach enables the imaging of the morphology of the cell adhering on the surface and can significantly contribute to multimodal ECL imaging and bioassays with different luminescent systems

Corrosion behaviour of single layered ZrNxOy thin films in artificial sweat solutions

Applications of coloured thin films can be found on the production of high-quality consumer products, such as eyeglass frames, wristwatch casings and wristbands. These components should possess scratch and corrosion resistant surfaces through the desired lifetime. Recently, metal oxynitrides, MeNxOy (Me = early transition metal) were proposed for decorative applications. In these materials, variations on the amount of oxygen allow the film properties to be tailored, originating a wide range of colours. Additionally, these materials should also fulfil the wear and corrosion requirements above referred. In the present work the corrosion behaviour of single layered zirconium oxynitride, ZrNxOy films, immersed in artificial sweat solutions, is described. Films were produced by rf reactive magnetron sputtering at a constant substrate temperature of 300 ºC, from a pure Zr target. The main processing variable was the flow rate of reactive. The corrosion resistance was evaluated by potentiodynamic polarisation tests and Electrochemical Impedance Spectroscopy (EIS) at different immersion times, at room temperature. The corrosion resistance of the films is strongly affected by the O/N ratio. A slight tendency to improving the corrosion resistance of the films was found with the increasing in the atomic fraction of oxygen. Nevertheless, pitting was found in all samples. However, the amount of pits seems to be strongly dependent not only on the composition of the film, but also on the processing-induced defects distribution.European Union, FCT- Portugal and European community (FEDER)

Tribocorrosion behaviour of zrNxOy thin films for decorative applications

The main aim of this work is the investigation of the tribocorrosion behaviour of single layered zirconium oxynitride, ZrNxOy, thin films in alternative linear regime of sliding and immersed in an artificial sweat solution at room temperature. The films were produced by rf reactive magnetron sputtering, using a pure Zr target at a constant temperature of 300º C. Two different sets of samples were produced. In the first set of films the substrate bias voltage was the main variable, whereas in the second set, the flow rate of reactive gases (oxygen/nitrogen ratio) was varied. The control of the amount of oxygen allowed the film properties to be tailored from those of covalent zirconium nitride to those of the correspondent ionic oxide. During the wear test both the open circuit potential and the corrosion current were monitored. Also, Electrochemical Impedance Spectroscopy (EIS) tests were performed before and after sliding in order to evaluate, in detail, the modification of the protective character of the coating introduced by the joint action of wear and corrosion. The modifications of the coating microstructure and/or chemical composition induced by the variation of the deposition parameters was also evaluated and correlated with the corrosion mechanisms occurring in each system

Corrosion behaviour of substoichiometric TiNx films produced by DC magnetron sputtering

The present work describes the corrosion behaviour of substoichiometric TiNx films obtained by dc reactive magnetron sputtering. The coatings thickness ranged from 1.7 to 4.2 µm and the nitrogen content varied between 0 and 55 at. %. According to structural characterization by XRD, the films revealed a hexagonal α-Ti phase with a strong [002] orientation for low nitrogen contents. For nitrogen contents of 20% and 30%, the ε-Ti2N phase appears with a [200] orientation and further increasing of nitrogen content showed that the δ-TiN phase was dominant. Potentiodynamic polarisation and Electrochemical Impedance Spectroscopy (EIS) techniques were used to study the corrosion properties of TiNx films when immersed in artificial sweat solutions. Results of potentiodynamic polarisation tests showed that all films have a high corrosion resistance reflected by corrosion current densities values lower than 0.7µA/cm2. Also, EIS tests corroborated the results obtained in the polarisation tests, showing that films containing low percentages of nitrogen (less than 8 %) reveal the best corrosion resistance. Further increases in nitrogen content lead to a decrease in corrosion resistance. An exception to this behaviour was found for the film, with 30 % N. This sample presents an excellent corrosion resistance which increases with the immersion time. Higher nitrogen contents (52 and 55 %) promote a relative increase in the corrosion resistance when compared with 45 and 50 at % films, but never reaching values obtained for nitrogen contents lower than 30 % at.Fundação para a Ciência e a Tecnologia - (FCT