Prediction of Response to Cisplatin-Based Neoadjuvant Chemotherapy of Muscle-Invasive Bladder Cancer Patients by Molecular Subtyping including KRT and FGFR Target Gene Assessment

Patients with muscle-invasive urothelial carcinoma achieving pathological complete response (pCR) upon neoadjuvant chemotherapy (NAC) have improved prognosis. Molecular subtypes of bladder cancer differ markedly regarding sensitivity to cisplatin-based chemotherapy and harbor FGFR treatment targets to various content. The objective of the present study was to evaluate whether preoperative assessment of molecular subtype as well as FGFR target gene expression is predictive for therapeutic outcome—rate of ypT0 status—to justify subsequent prospective validation within the “BladderBRIDGister”. Formalin-fixed paraffin-embedded (FFPE) tissue specimens from transurethral bladder tumor resections (TUR) prior to neoadjuvant chemotherapy and corresponding radical cystectomy samples after chemotherapy of 36 patients were retrospectively collected. RNA from FFPE tissues were extracted by commercial kits, Relative gene expression of subtyping markers (e.g., KRT5, KRT20) and target genes (FGFR1, FGFR3) was analyzed by standardized RT-qPCR systems (STRATIFYER Molecular Pathology GmbH, Cologne). Spearman correlation, Kruskal–Wallis, Mann–Whitney and sensitivity/specificity tests were performed by JMP 9.0.0 (SAS software). The neoadjuvant cohort consisted of 36 patients (median age: 69, male 83% vs. female 17%) with 92% of patients being node-negative during radical cystectomy after 1 to 4 cycles of NAC. When comparing pretreatment with post-treatment samples, the median expression of KRT20 dropped most significantly from DCT 37.38 to 30.65, which compares with a 128-fold decrease. The reduction in gene expression was modest for other luminal marker genes (GATA3 6.8-fold, ERBB2 6.3-fold). In contrast, FGFR1 mRNA expression increased from 33.28 to 35.88 (~6.8-fold increase). Spearman correlation revealed positive association of pretreatment KRT20 mRNA levels with achieving pCR (r = 0.3072: p = 0.0684), whereas pretreatment FGFR1 mRNA was associated with resistance to chemotherapy (r = −0.6418: p < 0.0001). Hierarchical clustering identified luminal tumors of high KRT20 mRNA expression being associated with high pCR rate (10/16; 63%), while the double-negative subgroup with high FGFR1 expression did not respond with pCR (0/9; 0%). Molecular subtyping distinguishes patients with high probability of response from tumors as resistant to neoadjuvant chemotherapy. Targeting FGFR1 in less-differentiated bladder cancer subgroups may sensitize tumors for adopted treatments or subsequent chemotherapy.     

The Distinct Role of the HDL Receptor SR-BI in Cholesterol Homeostasis of Human Placental Arterial and Venous Endothelial Cells

As opposed to adults, high-density lipoprotein (HDL) is the main cholesterol carrying lipoprotein in fetal circulation. The major HDL receptor, scavenger receptor class B type I (SR-BI), contributes to local cholesterol homeostasis. Arterial endothelial cells (ECA) from human placenta are enriched with cholesterol compared to venous endothelial cells (ECV). Moreover, umbilical venous and arterial plasma cholesterol levels differ markedly. We tested the hypothesis that the uptake of HDL-cholesteryl esters differs between ECA and ECV because of the differential expression of SR-BI. We aimed to identify the key regulators underlying these differences and the functional consequences. Immunohistochemistry was used for visualization of SR-BI in situ. ECA and ECV were isolated from the chorionic plate of human placenta and used for RT-qPCR, Western Blot, and HDL uptake assays with ³H- and ¹²⁵I-labeled HDL. DNA was extracted for the methylation profiling of the SR-BI promoter. SR-BI regulation was studied by exposing ECA and ECV to differential oxygen concentrations or shear stress. Our results show elevated SR-BI expression and protein abundance in ECA compared to ECV in situ and in vitro. Immunohistochemistry demonstrated that SR-BI is mainly expressed on the apical side of placental endothelial cells in situ, allowing interaction with mature HDL circulating in the fetal blood. This was functionally linked to a higher increase of selective cholesterol ester uptake from fetal HDL in ECA than in ECV, and resulted in increased cholesterol availability in ECA. SR-BI expression on ECV tended to decrease with shear stress, which, together with heterogeneous immunostaining, suggests that SR-BI expression is locally regulated in the placental vasculature. In addition, hypomethylation of several CpG sites within the SR-BI promoter region might contribute to differential expression of SR-BI between chorionic arteries and veins. Therefore, SR-BI contributes to a local cholesterol homeostasis in ECA and ECV of the human feto-placental vasculature.     

Alternative Approach to Modeling of Nucleation and Remelting in Powder Bed Fusion Additive Manufacturing

In powder bed fusion (PBF) of metals, energy of a laser or electron beam is utilized to form near-net-shaped parts from a powder bed. A very promising application of PBF lies in the direct control of the resulting microstructure by adjusting process parameters. Especially the intentional tilting of grains in one certain direction offers a complete new field of activity for additively produced parts specially designed for a given load case. Nucleation is essential for utilization of this effect. Herein, an alternative approach for modeling nucleation in PBF of metals is introduced. The model is not intended to cover every physical effect of this extreme complex phenomenon. Based on observations reported from different researchers in the field of PBF, a heuristic approach is applied to consider new grains within a cellular automata (CA)-based crystal growth model for PBF. Utilizing the implicit capability of the CA to model competitive growth of grains, this approach does not require any additional computational capacities. The numerical calculations are carried out using a message–passing interface (MPI) parallelization on a high-performance computing (HPC) cluster located at the Regional Research Center Erlangen. The numerical results are validated by means of experiments and electron backscatter diffractometric measurements.     

Influence of Cu Addition and Microstructural Configuration on the Creep Resistance and Mechanical Properties of an Fe-Based α/α′/α″ Superalloy

Introducing Cu nanoparticles is an effective mechanism for strengthening and toughening Fe-based materials such as ultra-high-strength steels. Herein, the effect of Cu on the mechanical properties of a novel Fe-based α/α′/α″ superalloy is studied. Compared to a Cu-free reference alloy, nanoindentation reveals an increase in hardness, which was associated with the formation of Cu nanoparticles. Both alloys show room temperature (RT) compressive plastic strain at maximum stress greater than 8%, irrespective of the heat-treatment. At RT and at 750 °C, the Cu-containing alloy exhibits a slightly higher strength, but the heat treatment has a more significant impact: A configuration of α-matrix and intermetallic α′/α″-phases forming an interpenetrating network is superior to a state with isolated precipitates. This difference vanishes in monotonic creep experiments, and under the same conditions, the Cu-containing alloy exhibits a twice as high creep rate despite a slightly higher precipitate fraction. This is linked to a higher lattice misfit and faster-coarsening kinetics. Post-mortem transmission electron microscopy analysis of the creep-deformed specimens identifies dislocation bypass as the dominant deformation mechanism. However, the presence of <010>{110} dislocations in the interfacial networks and evidence of dislocation activity within α′/α″ precipitates suggest the occurrence of shearing events.     

Liquid‐selenium‐enhanced grain growth of nanoparticle precursor layers for CuInSe2 solar cell absorbers

Large‐grained CuInSe₂ absorber layers are synthesized using a non‐vacuum process based on nanoparticle ink precursors and selenization by rapid thermal processing (RTP). The use of hydroxide‐based particles in organic solvents allows for the conversion with elemental selenium without the need to employ explosive and/or toxic H₂ or H₂Se gasses. Lateral grain sizes up to 4 µm are obtained through a novel RTP route, overcoming the inherently high layer porosity for previous nanoparticle processes. Morphological and elemental characterization at interrupted selenization steps suggests that liquid selenium can play a beneficial role in promoting layer densification and grain growth. Long carrier collection lengths in CuInSe₂ enable notable conversion efficiencies, despite the low minority carrier lifetimes of below 1 ns. Record efficiencies up to 8.73% highlight the potential of low‐cost, non‐vacuum deposition of chalcopyrite absorber layers with safe and simple precursors and processing routes. Copyright © 2014 John Wiley & Sons, Ltd.     

Spin-Flip Raman Scattering on Electrons and Holes in Two-Dimensional (PEA)2PbI4 Perovskites

The class of Ruddlesden–Popper type (PEA)₂PbI₄ perovskites comprises 2D structures whose optical properties are determined by excitons with a large binding energy of about 260 meV. It complements the family of other 2D semiconductor materials by having the band structure typical for lead halide perovskites, that can be considered as inverted compared to conventional III–V and II–VI semiconductors. Accordingly, novel spin phenomena can be expected for them. Spin-flip Raman scattering is used here to measure the Zeeman splitting of electrons and holes in a magnetic field up to 10 T. From the recorded data, the electron and hole Landé factors (g-factors) are evaluated, their signs are determined, and their anisotropies are measured. The electron g-factor value changes from +2.11 out-of-plane to +2.50 in-plane, while the hole g-factor ranges between -0.13 and -0.51. The spin flips of the resident carriers are arranged via their interaction with photogenerated excitons. Also the double spin-flip process, where a resident electron and a resident hole interact with the same exciton, is observed showing a cumulative Raman shift. Dynamic nuclear spin polarization induced by spin-polarized holes is detected in corresponding changes of the hole Zeeman splitting. An Overhauser field of the polarized nuclei acting on the holes as large as 0.6 T can be achieved.     

Numerical and Experimental Investigation on the Self-Healing Potential of Interpenetrating Metal–Ceramic Composites

An interpenetrating metal ceramic composite (IMCC) has been investigated regarding the potential as well as the feasibility of self-healing. Triggered by heating, cracks in the damaged composite located mainly in the Al₂O₃ ceramic or at the interface could be filled and closed by the liquid AlSi10Mg metal alloy. This healing procedure promises to reduce stress concentrations at crack tips and to improve the mechanical properties compared to the predamaged composite. Two different numerical approaches have been introduced to investigate this assumption and the potential of self-healed IMCCs for a best case scenario: 1) A simple 2D model to analyze the reduction of stress concentrations in front of a crack tip within the ceramic due to healing and 2) a 3D model based on CT-scan reconstructed microstructures to study how macroscopic mechanical properties can be restored depending on the amount of predamage. Further, the self-healing approach has been investigated experimentally for the same composite. Despite the fact that experimental self-healing of the investigated IMCC is only moderately feasible so far, the study shows the great potential that can still be exploited in order to extend the service life time of IMCC engineering components.