Ti–Si–C thin films produced by magnetron sputtering : correlation between physical properties, mechanical properties and tribological behavior

Ti–Si–C thin films were deposited onto silicon, stainless steel and high-speed steel substrates by magnetron sputtering, using different chamber configurations. The composition of the produced films was obtained by Electron Probe Micro-Analysis (EPMA) and the structure by X-ray diffraction (XRD). The hardness and residual stresses were obtained by depth-sensing indentation and substrate deflection measurements (using Stoney’s equation), respectively. The tribological behavior of the produced films was studied by pin-on-disc. The increase of the concentration of non-metallic elements (carbon and silicon) caused significant changes in their properties. Structural analysis revealed the possibility of the coexistence of different phases in the prepared films, namely Ti metallic phase ( alpha-Ti or beta-Ti) in the films with higher Ti content. The coatings with highest carbon contents, exhibited mainly a sub-stoichiometric fcc NaCl TiC-type structure. These structural changes were also confirmed by resistivity measurements, whose values ranged from 10E3 Ohm/sq for low non-metal concentration, up to 10E6 Ohm /sq for the highest metalloid concentration. Astrong increase of hardness and residual stresses was observed with the increase of the non-metal concentration in the films. The hardness (H) values ranged between 11 and 27 GPa, with a clear dependence on both crystalline structure and composition features. Following the mechanical behavior, the tribological results showed similar trends, with both friction coefficients and wear revealing also a straight correlation with the composition and crystalline structure of the coatings

Optimization and thermal stability of TiAlN-Mo multilayers

In this work we focus on the optimization and thermal stability of nanocomposite TiAlN/Mo multilayers that were produced by reactive magnetron sputtering on high-speed steel substrates, with modulation periods below 5 nm. These multilayers were annealed between 600– 900 ºC for 1 h in a vacuum furnace. Preliminary X-ray diffraction results reveal that these coatings are very stable up to 900 ºC, since the multilayer chemical modulation is not severely affected. At intermediate annealing temperatures the modulation period decreases due to interdiffusion at the interface, resulting in a thicker interface between metal/nitride and hence decreasing the thickness of those layers.Portuguese FCT/MCES scientific program

ZrOxNy decorative thin films prepared by the reactive gas pulsing process

Zirconium oxynitride thin films were deposited by dc reactive magnetron sputtering. A zirconium metallic target was sputtered in an Ar + N2 + O2 atmosphere. Argon and nitrogen flow rates were maintained constant whereas oxygen was pulsed during the deposition, implementing the reactive gas pulsing process (RGPP). A constant pulsing period T = 3 s was used following an exponential periodic signal versus time. The introduction time of oxygen was systematically changed from 17% to 83% of the period. The RGPP allowed the synthesis of ZrOxNy films with tuneable metalloid concentrations adjusting the introduction time of the oxygen. Composition and structural variations associated with mechanical, optical and electrical properties exhibited a smooth transition, from metallic-like characteristics, typical of the fcc-ZrN phase, to semi-conducting behaviour corresponding to a mixture of orthorhombic-Zr3N4(O) and γ -Zr2ON2 crystalline phases.Fundação para a Ciência e Tecnologia (FCT) - PTDC/CTM/69362/2006

An economic insight into additive manufacturing system implementation

With an ever growing diffusion of Additive Manufacturing (AM) system in industrial and commercial level, as well as the direct and indirect dynamics which are being introduced resulting from its inclusion as a possible production technology on companies’ portfolio, the need to reconfigure production system and adapt the production strategy becomes even more relevant than before. There are several studies which have emphasized on the importance of a paradigm shift in order to exploit advantages of AM, not only considering changes within design and functionality of the product, but also concerning AM’s impact on the entire value chain (re)configuration. Thus, it is of crucial importance to take into consideration that for this shift to be feasible and manageable, there is a need to include both technical and managerial aspects of manufacturing. This work proposes an economic insight in order to provide a guideline for the proper evaluation of AM system implementation. © IFIP International Federation for Information Processing 2015

Effect of thermal treatments on the structure of MoNxOy thin films

Article in PressMoNxOy films were deposited on steel substrates by dc reactive magnetron sputtering. The depositions were carried out from a pure molybdenum target, varying the flow rate of reactive gases. X-ray diffraction (XRD) results revealed the occurrence of cubic MoNx and hexagonal (d-MoN) phases for the films with high nitrogen flow rates. The increase of oxygen content induces the decrease of the grain size of the molybdenum nitride crystallites. The thermal stability of a set of samples was studied in vacuum, for an annealing time of 1 h, for temperatures ranging from 500 to 800 C in 100 C steps. The results showed that pure molybdenum nitride films changed their structure from a meta-stable cubic MoN to hexagonal d-MoN and cubic g-Mo2N-type structures with increasing annealing temperatures. The samples with molybdenum nitride films evidenced a good thermal stability, but those with molybdenum oxynitride coatings showed a tendency to detach with the increase of the annealing temperature.Fundação para a Ciência e a Tecnologia (FCT) - POCTI/CTM/38086/200

Recognition of vitamin B metabolites by mucosal-associated invariant T cells

The mucosal-associated invariant T-cell antigen receptor (MAIT TCR) recognizes MR1 presenting vitamin B metabolites. Here we describe the structures of a human MAIT TCR in complex with human MR1 presenting a non-stimulatory ligand derived from folic acid and an agonist ligand derived from a riboflavin metabolite. For both vitamin B antigens, the MAIT TCR docks in a conserved manner above MR1, thus acting as an innate-like pattern recognition receptor. The invariant MAIT TCR a-chain usage is attributable to MR1-mediated interactions that prise open the MR1 cleft to allow contact with the vitamin B metabolite. Although the non-stimulatory antigen does not contact the MAIT TCR, the stimulatory antigen does. This results in a higher affinity of the MAIT TCR for a stimulatory antigen in comparison with a non-stimulatory antigen. We formally demonstrate a structural basis for MAIT TCR recognition of vitamin B metabolites, while illuminating how TCRs recognize microbial metabolic signatures

Properties of MoNxOy thin films as a function of N/O ratio

The main purpose of this work consists on the preparation of single layered molybdenum oxynitride, MoNxOy. The films were deposited on steel substrates by dc reactive magnetron sputtering. The depositions were carried out from a pure Mo target varying the flow rate of reactive gases, which allowed tune the crystallographic structure between insulating oxides and metallic nitrides and consequently electronic, mechanical and optical properties of the material. X-ray diffraction (XRD) results revealed the occurrence of molybdenum nitride for the films with low oxygen fraction: face-centred cubic phases (gama-Mo2N) for low nitrogen flow rate or cubic MoNx and hexagonal phase (delta-MoN) for high nitrogen flow rate. The increase of oxygen content induces an amorphization of the nitride phases and appearance of MoO3 phases. The increase of the oxygen fraction in the films induces also a high decrease in films hardness. Residual stresses revealed to be of compressive type, in the range of very few tenths of GPa to 2 GPa. All these results have been analysed and will be presented as a function of the deposition parameters, the chemical composition and the structure of the films.Fundação para a Ciência e a Tecnologia (FCT) – Pograma Operacional “Ciência, Tecnologia, Inovação” - POCTI/CTM/38086/2001.Comunidade Europeia (CE). Fundo Europeu de Desenvolvimento Regional (FEDER)

Logical Development of the Cell Ontology

<p>Abstract</p> <p>Background</p> <p>The Cell Ontology (CL) is an ontology for the representation of <it>in vivo </it>cell types. As biological ontologies such as the CL grow in complexity, they become increasingly difficult to use and maintain. By making the information in the ontology computable, we can use automated reasoners to detect errors and assist with classification. Here we report on the generation of computable definitions for the hematopoietic cell types in the CL.</p> <p>Results</p> <p>Computable definitions for over 340 CL classes have been created using a genus-differentia approach. These define cell types according to multiple axes of classification such as the protein complexes found on the surface of a cell type, the biological processes participated in by a cell type, or the phenotypic characteristics associated with a cell type. We employed automated reasoners to verify the ontology and to reveal mistakes in manual curation. The implementation of this process exposed areas in the ontology where new cell type classes were needed to accommodate species-specific expression of cellular markers. Our use of reasoners also inferred new relationships within the CL, and between the CL and the contributing ontologies. This restructured ontology can be used to identify immune cells by flow cytometry, supports sophisticated biological queries involving cells, and helps generate new hypotheses about cell function based on similarities to other cell types.</p> <p>Conclusion</p> <p>Use of computable definitions enhances the development of the CL and supports the interoperability of OBO ontologies.</p

Endosomal MR1 Trafficking Plays a Key Role in Presentation of Mycobacterium tuberculosis Ligands to MAIT Cells

Mucosal-Associated Invariant T (MAIT) cells, present in high frequency in airway and other mucosal tissues, have Th1 effector capacity positioning them to play a critical role in the early immune response to intracellular pathogens, including Mycobacterium tuberculosis (Mtb). MR1 is a highly conserved Class I-like molecule that presents vitamin B metabolites to MAIT cells. The mechanisms for loading these ubiquitous small molecules are likely to be tightly regulated to prevent inappropriate MAIT cell activation. To define the intracellular localization of MR1, we analyzed the distribution of an MR1-GFP fusion protein in antigen presenting cells. We found that MR1 localized to endosomes and was translocated to the cell surface upon addition of 6-formyl pterin (6-FP). To understand the mechanisms by which MR1 antigens are presented, we used a lentiviral shRNA screen to identify trafficking molecules that are required for the presentation of Mtb antigen to HLA-diverse T cells. We identified Stx18, VAMP4, and Rab6 as trafficking molecules regulating MR1-dependent MAIT cell recognition of Mtb-infected cells. Stx18 but not VAMP4 or Rab6 knockdown also resulted in decreased 6-FP-dependent surface translocation of MR1 suggesting distinct pathways for loading of exogenous ligands and intracellular mycobacterially-derived ligands. We postulate that endosome-mediated trafficking of MR1 allows for selective sampling of the intracellular environment.Career Development Award: (#IK2 CX000538); U.S. Department of Veterans Affairs Clinical Sciences Research and Development Program (MJH); U.S.Department of Veterans Affairs Biomedical Laboratory Research and Development Program (DML) Merit Award: (#I01 BX000533); American Lung Association: (RT-350058)

A stable explant culture of HER2/neu invasive carcinoma supported by alpha-Smooth Muscle Actin expressing stromal cells to evaluate therapeutic agents

<p>Abstract</p> <p>Background</p> <p>To gain a better understanding of the effects of therapeutic agents on the tumor microenvironment in invasive cancers, we developed a co-culture model from an invasive lobular carcinoma. Tumor cells expressing HER2/neu organize in nests surrounded by alpha-Smooth Muscle Actin (α-SMA) expressing tumor stroma to resemble the morphology of an invading tumor. This co-culture, Mammary Adenocarcinoma Model (MAM-1) maintains a 1:1 ratio of HER2/neu positive tumor cells to α-SMA-reactive stromal cells and renews this configuration for over 20 passages in vitro.</p> <p>Methods</p> <p>We characterized the cellular elements of the MAM-1 model by microarray analysis, and immunocytochemistry. We developed flow cytometric assays to evaluate the relative responses of the tumor and stroma to the tyrosine kinase inhibitor, Iressa.</p> <p>Results</p> <p>The MAM-1 gene expression profile contains clusters that represent the ErbB-2 breast cancer signature and stroma-specific clusters associated with invasive breast cancers. The stability of this model and the ability to antigenically label the tumor and stromal fractions allowed us to determine the specificity of Iressa, a receptor tyrosine kinase inhibitor, for targeting the tumor cell population. Treatment resulted in a selective dose-dependent reduction in phospho-pMEK1/2 and pp44/42MAPK in tumor cells. Within 24 h the tumor cell fraction was reduced 1.9-fold while the stromal cell fraction increased >3-fold, consistent with specific reductions in phospho-pp44/42 MAPK, MEK1/2 and PCNA in tumor cells and reciprocal increases in the stromal cells. Erosion of the tumor cell nests and augmented growth of the stromal cells resembled a fibrotic response.</p> <p>Conclusion</p> <p>This model demonstrates the specificity of Iressa for HER2/neu expressing tumor cells versus the tumor associated myofibroblasts and is appropriate for delineating effects of therapy on signal transduction in the breast tumor microenvironment and improving strategies that can dually or differentially target the tumor and stromal elements in the microenvironment.</p