Sequential decisions on FAS sequencing guided by biomarkers in patients with lymphoproliferation and autoimmune cytopenia.

Clinical and genetic heterogeneity renders confirmation or exclusion of autoimmune lymphoproliferative syndrome difficult. To re-evaluate and improve the currently suggested diagnostic approach to patients with suspected FAS mutation, the most frequent cause of autoimmune lymphoproliferative syndrome, we prospectively determined 11 biomarkers in 163 patients with splenomegaly or lymphadenopathy and presumed or proven autoimmune cytopenia(s). Among 98 patients sequenced for FAS mutations in CD3(+)TCRα/ß(+)CD4(-)CD8(-) "double negative" T cells, 32 had germline and six had somatic FAS mutations. The best a priori predictor of FAS mutations was the combination of vitamin B12 and soluble FAS ligand (cut-offs 1255 pg/mL and 559 pg/mL, respectively), which had a positive predictive value of 92% and a negative predictive value of 97%. We used these data to develop a web-based probability calculator for FAS mutations using the three most discriminatory biomarkers (vitamin B12, soluble FAS ligand, interleukin-10) of the 11 tested. Since more than 60% of patients with lymphoproliferation and autoimmune cytopenia(s) in our cohort did not harbor FAS mutations, 15% had somatic FAS mutations, and the predictive value of double-negative T-cell values was rather low (positive and negative predictive values of 61% and 77%, respectively), we argue that the previously suggested diagnostic algorithm based on determination of double-negative T cells and germline FAS sequencing, followed by biomarker analysis, is not efficient. We propose vitamin B12 and soluble FAS ligand assessment as the initial diagnostic step with subsequent decision on FAS sequencing supported by a probability-calculating tool.

Reassessment of the Matrix Composition of Co-Re-Cr-Based Alloys for Particle Strengthening in High-Temperature Applications and Investigation of Suitable MC-Carbides.

This study reassesses the optimum matrix composition of Co-Re-Cr-based alloys for strengthening by MC-type carbides. It is found that the composition of Co-15Re-5Cr is ideally suited for this purpose as it allows the solution of the carbide-forming elements such as Ta, Ti, Hf, and C within a matrix consisting entirely of fcc-phase (typically at 1450 °C), having a high solubility for these elements, while precipitation heat treatment (typically at 900-1100 °C) occurs in a hcp-Co matrix, displaying a much lower solubility. In the case of the monocarbides TiC and HfC, this was investigated and achieved for the first time in Co-Re-based alloys. TaC and TiC emerged as suitable particles in Co-Re-Cr alloys for creep applications due to a large population of nano-sized particle precipitation, which is not the case for the mainly coarse HfC. Both Co-15Re-5Cr-xTa-xC and Co-15Re-5Cr-xTi-xC exhibit a formerly unknown maximum solubility close to x = 1.8 at.%. Therefore, further research on the particle-strengthening effect and the governing creep mechanisms of carbide-strengthened Co-Re-Cr alloys should focus on alloys with the following compositions: Co-15Re-5Cr-1.8Ta-1.8C and Co-15Re-5Cr-1.8Ti-1.8C.

Laser powder bed fusion (LPBF) of commercially pure titanium and alloy development for the LPBF process.

Laser powder bed fusion (LPBF) of titanium or titanium alloys allows fabrication of geometrically more complex and, possibly, individualized implants or osteosynthesis products and could thus improve the outcome of medical treatments considerably. However, insufficient LPBF process parameters can result in substantial porosity, decreasing mechanical properties and requiring post-treatment. Furthermore, texturized parts with anisotropic properties are usually obtained after LPBF processing, limiting their usage in medical applications. The present study addresses both: first, a design of experiments is used in order to establish a set of optimized process parameters and a process window for LPBF printing of small commercially pure (CP) titanium parts with minimized volume porosity. Afterward, the first results on the development of a biocompatible titanium alloy designed for LPBF processing of medical implants with improved solidification and more isotropic properties are presented on the basis of conventionally melted alloys. This development was performed on the basis of Ti-0.44O-0.5Fe-0.08C-0.4Si-0.1Au, a near-α alloy presented by the authors for medical applications and conventional manufacturing, with yttrium and boron additions as additional growth restriction solutes. In terms of LPBF processing of CP titanium grade 1 powder, a high relative density of approximately 99.9% was obtained in the as-printed state of the volume of a small cubical sample by using optimized laser power, scanning speed, and hatch distance in combination with a rotating scanning pattern. Moreover, tensile specimens processed with these volume settings and tested in the as-printed milled state exhibited a high average yield and ultimate tensile strength of approximately 663 and 747 N/mm2, respectively, combined with a high average ductility of approximately 24%. X-ray diffraction results suggest anisotropic mechanical properties, which are, however, less pronounced in terms of the tested specimens. Regarding alloy development, the results show that yttrium additions lead to a considerable microstructure refinement but have to be limited due to the occurrence of a large amount of precipitations and a supposed higher propensity for the formation of long columnar prior ß-grains. However, phase/texture and microstructure analyses indicate that Ti-0.44O-0.5Fe-0.08C-0.4Si-0.1Au-0.1B-0.1Y is a promising candidate to achieve lower anisotropy during LPBF processing, but further investigations on LPBF printing and Y2O3 formation are necessary.

New Approach in the Determination of a Suitable Directionally Coarsened Microstructure for the Fabrication of Nanoporous Superalloy Membranes Based on CMSX-4.

The pore size of nanoporous superalloy membranes produced by directional coarsening is directly related to the γ-channel width after creep deformation, since the γ-phase is removed subsequently by selective phase extraction. The continuous network of the γ'-phase thus remaining is based on complete crosslinking of the γ'-phase in the directionally coarsened state forming the subsequent membrane. In order to be able to achieve the smallest possible droplet size in the later application in premix membrane emulsification, a central aspect of this investigation is to minimize the γ-channel width. For this purpose, we use the 3w0-criterion as a starting point and gradually increase the creep duration at constant stress and temperature. Stepped specimens with three different stress levels are used as creep specimens. Subsequently, the relevant characteristic values of the directionally coarsened microstructure are determined and evaluated using the line intersection method. We show that the approximation of an optimal creep duration via the 3w0-criterion is reasonable and that coarsening occurs at different rates in dendritic and interdendritic regions. The use of staged creep specimens shows significant material and time savings in determining the optimal microstructure. Optimization of the creep parameters results in a γ-channel width of 119 ± 43 nm in dendritic and 150 ± 66 nm in interdendritic regions while maintaining complete crosslinking. Furthermore, our investigations show that unfavorable stress and temperature combinations favor undirectional coarsening before the rafting process is completed.

Investigation of TaC and TiC for Particle Strengthening of Co-Re-Based Alloys.

Cobalt-Rhenium (Co-Re)-based alloys are currently investigated as potential high-temperature materials with melting temperatures beyond those of nickel-based superalloys. Their attraction stems from the binary Co-Re phase diagram, exhibiting complete miscibility between Co and Re, whereby the melting temperature steadily increases with the Re-content. Thus, depending on the Re-content, one can tune the melting temperature between that of pure Co (1495 °C) and that of pure Re (3186 °C). Current investigations focus on Re-contents of about 15 at.%, which makes melting with standard equipment still feasible. In addition to solid solution strengthening due to the mixture of Co- and Re-atoms, particle strengthening by tantalum carbide (TaC) and titanium carbide (TiC) precipitates turned out to be promising in recent studies. Yet, it is currently unclear which of the two particle types is the best choice for high temperature applications nor has the strengthening mechanism associated with the monocarbide (MC)-precipitates been elucidated. To address these issues, we perform compression tests at ambient and elevated temperatures on the particle-free base material containing 15 at.% of rhenium (Re), 5 at.% of chromium (Cr) and cobalt (Co) as balance (Co-15Re-5Cr), as well as on TaC- and TiC-containing variants. Additionally, transmission electron microscopy is used to analyze the shape of the precipitates and their orientation relationship to the matrix. Based on these investigations, we show that TiC and TaC are equally suited for precipitation strengthening of Co-Re-based alloys and identify climb over the elongated particles as a rate controlling particle strengthening mechanism at elevated temperatures. Furthermore, we show that the Re-atoms are remarkably strong obstacles to dislocation motion, which are overcome by thermal activation at elevated temperatures.

Reducing the γ'-Particle Size in CMSX-4 for Membrane Development.

Colloidal emulsions for lipophilic drugs can be fabricated using premix membrane emulsification. The state of the art is the application of membranes made from, for example, polycarbonate or polyester, which, however, are prone to fouling and cause waste, due to the low number of cycles. With the use of metallic membranes made from the nickel based single crystalline superalloy CMSX-4, these key disadvantages are eliminated. However, instead, the pore size and the resulting droplet size distribution need to be adjusted and improved. This can be realized by tailoring the size of the γ'-particles, which is controllable by the time and temperature used during precipitation heat treatment and the quenching method after homogenization heat treatment. Therefore, we utilized different heat treatment protocols, varying the cooling rate (water quenching and air cooling) after homogenization heat treatment and the holding time and temperature during precipitation heat treatment. Then, we investigated the γ/γ'-microstructure, including the γ'-morphology and γ'-particle size. We show that water quenching has a significant impact on the γ/γ'-microstructure and often leads to irregular-shaped and poorly aligned γ'-particles after precipitation heat treatment. In comparison, air cooling, followed by a subsequent precipitation heat treatment, results in well-aligned and cubic shaped γ'-particles and is, therefore, favorable for membrane fabrication. A reduction in precipitation temperature leads to morphology changes to the γ'-particles. A reduction of the holding time during precipitation heat treatment diminishes the γ'-particle growth, resulting in smaller γ'-particles. Additionally, a suitable heat treatment protocol for membrane fabrication was identified with a γ'-edge length of 224 ± 52 nm and well-aligned, cubic shaped γ'-particles.

Polycrystalline Superalloy Membranes Produced by Load-Free Coarsening of Incoherent γ'-Precipitates: Microstructure Evolution and Mechanical Properties.

Nanoporous superalloy membranes are a functional extension of the use of nickel-based alloys. The material, which is usually used for high-temperature applications, consists mainly of the two phases γ and γ'. Through coarsening of the precipitates and thus forming of a bicontinuous γ/γ' network, membranes can be produced by removing either of these phases. From the single-crystalline alloy CMSX-4, the bicontinuous network can be formed either thermo-mechanically by directional coarsening of coherent precipitates or by load-free coalescence of incoherent precipitates. Recent investigations have shown that membranes also can be produced from polycrystalline starting material in both ways. In this article, the process route for membranes by load-free coarsening of incoherent γ' precipitates from a carbon-free version of the polycrystalline alloy Nimonic 115 is presented. This manufacturing method has the advantage of its simplicity and in comparison to single-crystalline membranes it can be realized in larger scales. We discuss the microstructure and show the mechanical properties by means of tensile tests. Despite the grain boundaries as a mechanical weak link, polycrystalline membranes show promising mechanical properties. Their strength even exceeds that of the single-crystalline membranes despite the significantly higher pore volume content.

Nanoporous membranes produced from polycrystalline Ni-based superalloys.

Porous metal membranes are produced from a two phase system in which the discrete cubic gamma'-precipitates connect during self assembly. In the so called rafting process the cubic particles start to coarsen and finally create a network within the gamma-matrix. In a following electrochemical leaching process one of the phases can be removed leaving the nanoporous membrane. So far, single crystalline alloys have been used for producing thin nanoporous membranes. Now research is in progress to produce the nanoporous membranes from polycrystalline alloys in a creep process. A modification of the commercially available alloy Nimonic 115 was used for these membranes. The permeability of these membranes was proven in a gas-flow test.

Identification of Material Parameters for the Simulation of Acoustic Absorption of Fouled Sintered Fiber Felts.

As a reaction to the increasing noise pollution, caused by the expansion of airports close to residential areas, porous trailing edges are investigated to reduce the aeroacoustic noise produced by flow around the airframe. Besides mechanical and acoustical investigations of porous materials, the fouling behavior of promising materials is an important aspect to estimate the performance in long-term use. For this study, two sintered fiber felts were selected for a long-term fouling experiment where the development of the flow resistivity and accumulation of dirt was observed. Based on 3D structural characterizations obtained from X-ray tomography of the initial materials, acoustic models (Biot and Johnson-Champoux-Allard) in the frame of the transfer matrix method were applied to the sintered fiber felts. Flow resistivity measurements and the measurements of the absorption coefficient in an impedance tube are the basis for a fouling model for sintered fiber felts. The contribution will conclude with recommendations concerning the modeling of pollution processes of porous materials.

Nanoforging - Innovation in three-dimensional processing and shaping of nanoscaled structures.

BACKGROUND: This paper describes the shaping of freestanding objects out of metallic structures in the nano- and submicron size. The technique used, called nanoforging, is very similar to the macroscopic forging process. RESULTS: With spring actuated tools produced by focused ion beam milling, controlled forging is demonstrated. With only three steps, a conical bar stock is transformed to a flat- and semicircular bent bar stock. CONCLUSION: Compared with other forming techniques in the reduced scale, nanoforging represents a beneficial approach in forming freestanding metallic structures, due to its simplicity, and supplements other forming techniques.

Cytocompatibility of a free machining titanium alloy containing lanthanum.

Titanium alloys like Ti6Al4V are widely used in medical engineering. However, the mechanical and chemical properties of titanium alloys lead to poor machinability, resulting in high production costs of medical products. To improve the machinability of Ti6Al4V, 0.9% of the rare earth element lanthanum (La) was added. The microstructure, the mechanical, and the corrosion properties were determined. Lanthanum containing alloys exhibited discrete particles of cubic lanthanum. The mechanical properties and corrosion resistance were slightly decreased but are still sufficient for many applications in the field of medical engineering. In vitro experiments with mouse macrophages (RAW 264.7) and human bone-derived cells (MG-63, HBDC) were performed and revealed that macrophages showed a dose response below and above a LaCl3 concentration of 200 microM, while MG-63 and HBDC tolerated three times higher concentrations without reduction of viability. The viability of cells cultured on disks of the materials showed no differences between the reference and the lanthanum containing alloy. We therefore propose that lanthanum containing alloy appears to be a good alternative for biomedical applications, where machining of parts is necessary.

T cell responses to hepatitis B surface antigen are detectable in non-vaccinated individuals.

AIM: To evaluate, whether humoral hepatitis-B-vaccine non-responders also fail to mount a T cell response and to compare these results to normal vaccinees. METHODS: Fourty-seven health care employees were enrolled in this study including all available non-responders (n = 13) with an anti-HBsAg titer < 10 kU/L and all available low-responders (n = 12) with an anti-HBsAg titer < 100 kU/L. Also, 12 consecutive anti-HBsAg negative pre-vaccination subjects were enrolled as well as 10 subjects (+7 from the vaccinated group) with titers > 1000 kU/L as controls. PBMC from all subjects were analyzed by IFN-gamma and IL-4 ELISPOT assays for the presence of hepatitis B surface antigen (HBsAg) reactive T cells. RESULTS: Non-responders and low-responders had no or only very limited T cell responses, respectively. Individuals responding to vaccination with the induction of a high anti-HBsAg titer showed a strong T cell response after the third vaccination. Surprisingly, these individuals showed response even before the first vaccination. T cell response to control antigens and mitogens was similar in all groups. CONCLUSION: Our data suggest that there is no general immune deficiency in non-/low-responders. Thus, we hypothesize that the induction of anti-HBsAg responses by vaccination is significantly dependent on the pre-existing T cell repertoire against the specific antigen rather than the presence of a general T cell defect.