Evaluation of miRNA detection methods for the analytical characteristics necessary for clinical utilization

miRNAs are promising biomarkers but methods for their measurement are not clear. We therefore examined three miRNA detection technologies and considered the analytical characteristics essential for clinical utilization. TaqMan assays, SplintR-qPCR and miREIA were compared for their absolute quantification bias, conformity and robustness. Absolute concentrations of miR-142-5p, miR-23a-3p and miR-93-5p were measured with all three methods using 30 samples. Robustness was evaluated by measurement of miR-21-5p in five uniform experiments. Correlations were miRNA-specific, but we observed a different absolute concentration range in RT-qPCR (fmol/mu l) and methods evading the RT process (amol/mu l). Consistently, RT-less methods reported better robustness (CV 8-19%) than RT-qPCR (CV 39-50%). The calibration curve in TaqMan Advanced assay was influenced by dilution media. Methods avoiding RT seem to be a promising future alternative for miRNA measurement. METHOD SUMMARY Three miRNA detection technologies were compared: 1) RT-qPCR where the RT step was performed with either a specific (TaqMan miRNA assay) or universal (TaqMan Advanced assay) priming strategy; 2) miREIA technology, using hybridization and specific antibody to DNA/RNA hybrids and 3) SplintR-qPCR, which utilizes a hybridization and ligation step followed by qPCR

Evolution of the Fe-Rich Phases in Recycled AlSi9Cu3 Cast Alloy during Solution Treatment

Using recycled aluminium cast alloys is profitable in many aspects. Requiring only 5% of the energy to produce secondary metal as compared to primary metal and generates only 4% of the CO2 emissions, the recycling of aluminium is therefore beneficial of both environmental and economical point of view. AlSi9Cu3 is a very frequently employed die-casting alloy. It is used in almost all fields of pressure die casting, especially in the fabrication various motor mounts, pistons, cylinder heads, heat exchangers, air conditioners or gearings. Al-Si-Cu alloys usually contain Cu and sometimes Mg as the main alloying elements, together with various impurities such as Fe, Mn or Cr. Presence of the Fe impurity enables the crystallization of brittles needles Al5FeSi during solidification, even thought the Fe content is extremely low in a melt, and its presence severely lowers the dynamic fracture toughness of the alloy. Alloying elements such are Mn, Cr, Mo and Be have beenused to change the morphology of the Al5 FeSi needles and Al(FeMnMg)Si skeleton (or "Chinese script") phases.The present study is a part of larger research project, which was conducted to investigate and to provide a better understanding of the solution annealing on the morphology of Fe-rich intermetallic phases in cast Al-Si-Cu alloys. The Fe-rich changes of typical specimens under different solution heat treatment condition were examined using an optical and scanning electron microscope (SEM). The results show that optimal solution treatment causes increasing of mechanical properties, the change of Si-morphology and reduction iron rich phases

Mechanical Properties of Heat Treated Secondary AlSi12Cu1Fe Cast Alloy at Room Temperature

The contribution describes changes of mechanical properties (strength tensile, absorbed energy and Brinell hardness) in secondary (recycled) eutectic aluminium-silicon cast alloy - AlSi12Cu1Fe during solution treatment. This work presents the influences of the solution treatment by 525 °C, 545 °C and 565 °C with the holding time 2, 4, 8, 16 and 32 hours, water quenching at 40 °C and natural aging with holding time 24 hours. Mechanical properties were measured in line with STN EN ISO at room temperature. Solution treatment has led to changes in mechanical properties, caused by the changes in microstructure including the spheroidization and coarsening of eutectic silicon, gradual disintegration, shortening and thinning of intermetallic phases