Digital PCR Panel for Sensitive Hematopoietic Chimerism Quantification after Allogeneic Stem Cell Transplantation

Accurate and sensitive determination of hematopoietic chimerism is a crucial diagnostic measure after allogeneic stem cell transplantation to monitor engraftment and potentially residual disease. Short tandem repeat (STR) amplification, the current “gold standard” for chimerism assessment facilitates reliable accuracy, but is hampered by its limited sensitivity (≥1%). Digital PCR (dPCR) has been shown to combine exact quantification and high reproducibility over a very wide measurement range with excellent sensitivity (routinely ≤0.1%) and thus represents a promising alternative to STR analysis. We here aimed at developing a whole panel of digital-PCR based assays for routine diagnostic. To this end, we tested suitability of 52 deletion/insertion polymorphisms (DIPs) for duplex analysis in combination with either a reference gene or a Y-chromosome specific PCR. Twenty-nine DIPs with high power of discrimination and good performance were identified, optimized and technically validated. We tested the newly established assays on retrospective patient samples that were in parallel also measured by STR amplification and found excellent correlation. Finally, a screening plate for initial genotyping with DIP-specific duplex dPCR assays was designed for convenient assay selection. In conclusion, we have established a comprehensive dPCR system for precise and high-sensitivity measurement of hematopoietic chimerism, which should be highly useful for clinical routine diagnostics.     

Mechanical and thermal characterization of as-received recycled polyethylene filled with rice husk and their relationship to the end use of these composites

ABSTRACT

Waste can be considered a crisis across the world, especially in the Global South where landfills are collapsing and vector-borne illnesses are increasing. Due to the increase in the amount of waste plastic in the environment, different uses of recycled plastics are being investigated. One such use is roofing tiles. This has been put in place in Sri Lanka through the Waste for Life organization. Here, recycled polyethylene is filled with rice husk particles. Due to the hydrophilic nature of the rice husk, the strength of the polyethylene composites decreased with increasing rice husk filler. Since the roofing tiles do not need to support a large load, the loss in strength should not be too detrimental. However, the creep behavior was improved with 10% and 20% filler. This is important in that the temperature on the roof can reach high temperatures.     

The effect of amorphous chalcogenides on mechanical and anticorrosive properties of protective organic coatings containing high amounts of zinc metal particles

The objective of this paper concerns reduction of zinc metal content in organic coatings while preserving their high anticorrosive efficiency. The two goals can be achieved by using amorphous chalcogenides as components of the protective coating. Special attention was paid to materials containing Ge₂₀Se₈₀, Ge₃₀Se₇₀ and Ge₄₀Se₆₀, which were characterized by physico-chemical properties. An epoxy ester resin was used as binder for the investigated organic coatings. Organic coatings were prepared by combining zinc metal with amorphous chalcogenides. The resistance of the prepared films was evaluated using the results of mechanical tests. The anticorrosive efficiency of the prepared films was evaluated using the results of direct corrosion tests. Thus the following conclusion can be made from the results of anti-corrosive tests: the higher the pigment volume concentration of amorphous chalcogenides in the coatings, the higher the protective performance of the paint against corrosion.     

Using the Default Option Bias to Influence Decision Making While Driving

Gaining a better understanding of human–computer interaction in multiple-goal environments, such as driving, is critical as people increasingly use information technology to accomplish multiple tasks simultaneously. Extensive research shows that decision biases can be utilized as effective cues to guide user interaction in single-goal environments. This article is a first step toward understanding the effect of decision biases in multiple-goal environments. This study analyzed data from a field experiment during which a comparison was made between drivers’ decisions on parking lots in a single-goal environment and drivers’ decisions in a multiple-goal environment when being exposed to the default option bias. The article shows that the default option bias is effective in multiple-goal environments. The results have important implications for the design of human–computer interaction in multiple-goal environments.     

Variable Frequency Microwave Curing of Amide-Epoxy Based Polymers

Irradiation of a dielectric material with microwave irradiation results in energy transfer due to rotational-vibrational transitions within the molecule. The energy transfer results in a local rise in temperature within the material. Microwave-induced reactions can occur at a lower average temperature than convective heating resulting in faster thermal curing of polymer dielectrics because the energy absorption is localized at particular sites. In this study, variable frequency microwave (VFM) curing of epoxy-based dielectric films was investigated. The microwave energy was swept through a range of frequencies to dissipate standing waves so that metallic conductors could be present with the dielectric films. The rate of reaction and film properties of polyamideimide (PAI) and bisphenol A epoxy resin (BPAEp) were studied. Benzanilide and BPAEp were used as model compounds for the reaction between the amide and epoxy. Compared to convective heating, the microwave reaction rates were higher at each isothermal cure temperature. The resulting mechanical properties of the films cured by microwave heated were superior to thermally cured materials. The elastic modulus of VFM cured PAI/BPAEp films was less than that of thermally cured films and the elongation to break was twice as high. Further, the adhesion to copper was improved with microwave processing. The chemical structure of the VFM cured polymer was different from the thermally cured polymer, based on Fourier transform infrared analysis, and is likely the origin of the improved properties.     

Proteins Marking the Sequence of Genotoxic Signaling from Irradiated Mesenchymal Stromal Cells to CD34+ Cells

Non-targeted effects (NTE) of ionizing radiation may initiate myeloid neoplasms (MN). Here, protein mediators (I) in irradiated human mesenchymal stromal cells (MSC) as the NTE source, (II) in MSC conditioned supernatant and (III) in human bone marrow CD34+ cells undergoing genotoxic NTE were investigated. Healthy sublethal irradiated MSC showed significantly increased levels of reactive oxygen species. These cells responded by increasing intracellular abundance of proteins involved in proteasomal degradation, protein translation, cytoskeleton dynamics, nucleocytoplasmic shuttling, and those with antioxidant activity. Among the increased proteins were THY1 and GNA11/14, which are signaling proteins with hitherto unknown functions in the radiation response and NTE. In the corresponding MSC conditioned medium, the three chaperones GRP78, CALR, and PDIA3 were increased. Together with GPI, these were the only four altered proteins, which were associated with the observed genotoxic NTE. Healthy CD34+ cells cultured in MSC conditioned medium suffered from more than a six-fold increase in γH2AX focal staining, indicative for DNA double-strand breaks, as well as numerical and structural chromosomal aberrations within three days. At this stage, five proteins were altered, among them IQGAP1, HMGB1, and PA2G4, which are involved in malign development. In summary, our data provide novel insights into three sequential steps of genotoxic signaling from irradiated MSC to CD34+ cells, implicating that induced NTE might initiate the development of MN.     

IV. Referate

Ohne Zusammenfassung     

Fabrication of air-channel structures for microfluidic,microelectromechanical, and microelectronic applications

A method is presented for fabricating micro-air-channel structures encapsulated by a dielectric material using a sacrificial polymer based on polynorbornene (PNB) chemistry. A spin-coated film of PNB was patterned to define the exact geometry of the air-channels using conventional lithographic and etching techniques. The sacrificial polymer was encapsulated with a permanent dielectric material. The composite was then raised to elevated temperatures to produce gaseous products which permeate through the encapsulating material (SiO₂ , SiN_x or other polymer) leaving behind minimal solid residue. Air-channels integrated with metal interconnections can be formed via a Damascene, or in-lay process. After patterning the sacrificial polymer, copper was electroplated, followed by encapsulation with the dielectric. Various issues pertaining to the processing steps have been investigated and are discussed, such as type of encapsulants, feasible air-channel sizes, and processing conditions. Such air-channel structures are believed to have potential applications in microelectronics, displays, printers, multilevel wiring boards, microscale chemical reactors on a chip, and microelectromechanical devices     

Quantitative electron spectroscopic diffraction analyses of the crystal formation in dentine

Newly formed apatitic crystallites of different hard tissues consist, according to our investigations, of chains composed of nanometre-sized particles (islands, dots) arising at nucleating sites of the collagenous and noncollagenous matrix macromolecules. In dentine these islands coalesce rapidly in longitudinal direction to form needle-like crystallites which further coalesce to ribbon-like crystallites. We have concluded that the centre-to-centre distances between these islands represent the distances between the nucleating sites of the matrix macromolecules. We have applied energy-filtering transmission electron microscopy in the selected area electron diffraction mode at different stages of crystal formation in dentine and have obtained quantitative information of the degree of crystal disorder on the basis of the paracrystal theory. The fluctuation of the lattice plane distances in c -axis direction decreases, proceeding from the region near the dentine/predentine border to the dentine/enamel border.