Interaction Effects in a 1D Flat Band at a Topological Crystalline Step Edge.

Step edges of topological crystalline insulators can be viewed as predecessors of higher-order topology, as they embody one-dimensional edge channels embedded in an effective three-dimensional electronic vacuum emanating from the topological crystalline insulator. Using scanning tunneling microscopy and spectroscopy, we investigate the behavior of such edge channels in Pb1-xSnxSe under doping. Once the energy position of the step edge is brought close to the Fermi level, we observe the opening of a correlation gap. The experimental results are rationalized in terms of interaction effects which are enhanced since the electronic density is collapsed to a one-dimensional channel. This constitutes a unique system to study how topology and many-body electronic effects intertwine, which we model theoretically through a Hartree-Fock analysis.

Synthesis of large scale 3D microscopic images of 3D cell cultures for training and benchmarking.

The analysis of 3D microscopic cell culture images plays a vital role in the development of new therapeutics. While 3D cell cultures offer a greater similarity to the human organism than adherent cell cultures, they introduce new challenges for automatic evaluation, like increased heterogeneity. Deep learning algorithms are able to outperform conventional analysis methods in such conditions but require a large amount of training data. Due to data size and complexity, the manual annotation of 3D images to generate large datasets is a nearly impossible task. We therefore propose a pipeline that combines conventional simulation methods with deep-learning-based optimization to generate large 3D synthetic images of 3D cell cultures where the labels are known by design. The hybrid procedure helps to keep the generated image structures consistent with the underlying labels. A new approach and an additional measure are introduced to model and evaluate the reduced brightness and quality in deeper image regions. Our analyses show that the deep learning optimization step consistently improves the quality of the generated images. We could also demonstrate that a deep learning segmentation model trained with our synthetic data outperforms a classical segmentation method on real image data. The presented synthesis method allows selecting a segmentation model most suitable for the user's data, providing an ideal basis for further data analysis.

3D Melanoma Cocultures as Improved Models for Nanoparticle-Mediated Delivery of RNA to Tumors.

Cancer therapy is an emergent application for mRNA therapeutics. While in tumor immunotherapy, mRNA encoding for tumor-associated antigens is delivered to antigen-presenting cells in spleen and lymph nodes, other therapeutic options benefit from immediate delivery of mRNA nanomedicines directly to the tumor. However, tumor targeting of mRNA therapeutics is still a challenge, since, in addition to delivery of the cargo to the tumor, specifics of the targeted cell type as well as its interplay with the tumor microenvironment are crucial for successful intervention. This study investigated lipoplex nanoparticle-mediated mRNA delivery to spheroid cell culture models of melanoma. Insights into cell-type specific targeting, non-cell-autonomous effects, and penetration capacity in tumor and stroma cells of the mRNA lipoplex nanoparticles were obtained. It was shown that both coculture of different cell types as well as three-dimensional cell growth characteristics can modulate distribution and transfection efficiency of mRNA lipoplex formulations. The results demonstrate that three-dimensional coculture spheroids can provide a valuable surplus of information in comparison to adherent cells. Thus, they may represent in vitro models with enhanced predictivity for the in vivo activity of cancer nanotherapeutics.

Life cycle inventory data generation by process simulation for conventional, feedstock recycling and power-to-X technologies for base chemical production.

The article presents the methodology and applicable data for the generation of life cycle inventory for conventional and alternative processes for base chemical production by process simulation. Addressed base chemicals include lower olefins, BTX aromatics, methanol, ammonia and hydrogen. Assessed processes include conventional chemical production processes from naphtha, LPG, natural gas and heavy fuel oil; feedstock recycling technologies via gasification and pyrolysis of refuse derived fuel; and power-to-X technologies from hydrogen and CO2. Further, process variations with additional hydrogen input are covered. Flowsheet simulation in Aspen Plus is applied to generate datasets with conclusive mass and energy balance under uniform modelling and assessment conditions with available validation data. Process inventory data is generated with no regard to the development stage of the respective technology, but applicable process data with high technology maturity is prioritized for model validation. The generated inventory data can be applied for life cycle assessments. Further, the presented modelling and balancing framework can be applied for inventory data generation of similar processes to ensure comparability in life cycle inventory data.

Global warming potential and economic performance of gasification-based chemical recycling and incineration pathways for residual municipal solid waste treatment in Germany.

Chemical recycling could facilitate the transition from a linear to a circular carbon economy, where carbon-containing waste is channeled back into the production cycle as a chemical feedstock instead of being incinerated or landfilled. However, the predominant focus on technological aspects of chemical recycling for plastic waste narrows evaluations of its potential in contributing to such a transition. Moreover, it leads to significant controversy about its role in the waste hierarchy as a possible competitor to mechanical recycling. To address these gaps in the literature, this study assesses ecological and economic impacts associated with chemical recycling of residual municipal solid waste in Germany. Combining approaches of life cycle assessment and techno-economic analysis, chemical recycling and conventional incineration-based treatment pathways are comparatively evaluated in terms of global warming potential and economic performance (i.e. fixed capital investment, net present value, dynamic payback period, and levelized cost of carbon abatement). Results indicate that compared to incineration-based conventional pathways, chemical recycling can contribute to reducing greenhouse gas emissions in low-emission energy systems. However, the economic performance of chemical recycling is highly dependent on its scale of operation. Additionally, a price premium for recycling products as well as economic instruments for penalizing CO2 emissions are identified to play important roles in the economic performance of chemical recycling.

Extracellular Matrix Components Regulate Bone Sialoprotein Expression in MDA-MB-231 Breast Cancer Cells.

Bone sialoprotein (BSP) has become a target in breast cancer research as it is associated with tumor progression and metastasis. The mechanisms underlying the regulation of BSP expression have been largely elusive. Given that BSP is involved in the homing of cancer cells in bone metastatic niches, we addressed regulatory effects of proteolytic cleavage and extracellular matrix components on BSP expression and distribution in cell culture models. Therefore, MDA-MB-231 human breast cancer cells were kept in 2D and 3D spheroid cultures and exposed to basement membrane extract in the presence or absence of matrix metalloproteinase 9 or the non-polar protease, dispase. Confocal imaging of immunofluorescence samples stained with different antibodies against human BSP demonstrated a strong inducing effect of basement membrane extract on anti-BSP immunofluorescence. Similarly, protease incubation led to acute upregulation of anti-BSP immunofluorescence signals, which was blocked by cycloheximide, suggesting de novo formation of BSP. In summary, our data show that extracellular matrix components play an important function in regulating BSP expression and hint at mechanisms for the formation of bone-associated metastasis in breast cancer that might involve local control of BSP levels by extracellular matrix degradation and release of growth factors.

A Scaffold-Free 3-D Co-Culture Mimics the Major Features of the Reverse Warburg Effect In Vitro.

Most tumors consume large amounts of glucose. Concepts to explain the mechanisms that mediate the achievement of this metabolic need have proposed a switch of the tumor mass to aerobic glycolysis. Depending on whether primarily tumor or stroma cells undergo such a commutation, the terms 'Warburg effect' or 'reverse Warburg effect' were coined to describe the underlying biological phenomena. However, current in vitro systems relying on 2-D culture, single cell-type spheroids, or basal-membrane extract (BME/Matrigel)-containing 3-D structures do not thoroughly reflect these processes. Here, we aimed to establish a BME/Matrigel-free 3-D microarray cancer model to recapitulate the metabolic interplay between cancer and stromal cells that allows mechanistic analyses and drug testing. Human HT-29 colon cancer and CCD-1137Sk fibroblast cells were used in mono- and co-cultures as 2-D monolayers, spheroids, and in a cell-chip format. Metabolic patterns were studied with immunofluorescence and confocal microscopy. In chip-based co-cultures, HT-29 cells showed facilitated 3-D growth and increased levels of hexokinase-2, TP53-induced glycolysis and apoptosis regulator (TIGAR), lactate dehydrogenase, and: translocase of outer mitochondrial membrane 20 (TOMM20), when compared with HT-29 mono-cultures. Fibroblasts co-cultured with HT-29 cells expressed higher levels of mono-carboxylate transporter 4, hexokinase-2, microtubule-associated proteins 1A/1B light chain 3, and ubiquitin-binding protein p62 than in fibroblast mono-cultures, in both 2-D cultures and chips. Tetramethylrhodamin-methylester (TMRM) live-cell imaging of chip co-cultures revealed a higher mitochondrial potential in cancer cells than in fibroblasts. The findings demonstrate a crosstalk between cancer cells and fibroblasts that affects cellular growth and metabolism. Chip-based 3-D co-cultures of cancer cells and fibroblasts mimicked features of the reverse Warburg effect.

Routine Optical Clearing of 3D-Cell Cultures: Simplicity Forward.

Three-dimensional cell cultures, such as spheroids and organoids, serve as increasingly important models in fundamental and applied research and start to be used for drug screening purposes. Optical tissue clearing procedures are employed to enhance visualization of fluorescence-stained organs, tissues, and three-dimensional cell cultures. To get a more systematic overview about the effects and applicability of optical tissue clearing on three-dimensional cell cultures, we compared six different clearing/embedding protocols on seven types of spheroid- and chip-based three-dimensional cell cultures of approximately 300 µm in size that were stained with nuclear dyes, immunofluorescence, cell trackers, and cyan fluorescent protein. Subsequent whole mount confocal microscopy and semi-automated image analysis were performed to quantify the effects. Quantitative analysis included fluorescence signal intensity and signal-to-noise ratio as a function of z-depth as well as segmentation and counting of nuclei and immunopositive cells. In general, these analyses revealed five key points, which largely confirmed current knowledge and were quantified in this study. First, there was a massive variability of effects of different clearing protocols on sample transparency and shrinkage as well as on dye quenching. Second, all tested clearing protocols worked more efficiently on samples prepared with one cell type than on co-cultures. Third, z-compensation was imperative to minimize variations in signal-to-noise ratio. Fourth, a combination of sample-inherent cell density, sample shrinkage, uniformity of signal-to-noise ratio, and image resolution had a strong impact on data segmentation, cell counts, and relative numbers of immunofluorescence-positive cells. Finally, considering all mentioned aspects and including a wish for simplicity and speed of protocols - in particular, for screening purposes - clearing with 88% Glycerol appeared to be the most promising option amongst the ones tested.

COVID-19: guaranteed Loans and Zombie Firms.

Based on a survey (7-13 April 2020) we evaluate the reaction of Swiss firms towards the COVID-19 crisis. Firms show little pro-active reactions towards the crisis, but decrease their business activities. The firms in the survey report that the decline in foreign demand is the single most important reason for their deteriorating business situation. Firms that faced a more difficult business situation before the crisis are affected more severely during the crisis. Moreover, we investigate the impact of the Swiss federal loan program (Bundeshilfe) on the business activities. To this end, we develop a stylized theoretical model of financially constrained heterogeneous firms. We find that policy makers face a trade-off between immediate higher unemployment rates and long-term higher public spending. The former arises from a combination of a too strong economic impact of the COVID-19 lockdown (demand drop) and too low levels of loans provided. Nevertheless, providing (too) high levels of loans to firms creates zombie firms that are going to default in the future leading to an increase in public spending. (JEL codes: D22, D25, D84, and G33).

Bone Sialoprotein Shows Enhanced Expression in Early, High-Proliferation Stages of Three-Dimensional Spheroid Cell Cultures of Breast Cancer Cell Line MDA-MB-231.

Normally, bone sialoprotein (BSP) is an important contributor to bone micro-calcification. However, it is also highly expressed in bone-metastatic malignancies, including prostate, lung, and breast cancer. In these disorders, BSP correlates with poor prognosis. Its expression in triple-negative breast cancer cells is enhanced by the transcription factor RUNX2, and both, BSP and RUNX2 are under control of IGF-1 and TGFß1. Knockdown of BSP or its inactivation by specific antibodies were found to reduce the metastatic potential of MDA-MB-231 triple-negative breast cancer cells in xenografts. While the role of BSP in bone metastasis was studied using such in vivo models, valid in vitro test systems to investigate BSP biology have been lacking since this protein is expressed at very low levels in classical 2D cell cultures and the frequently used breast cancer cell line MDA-MB-231 is difficult to grow in 3D. Here, we have developed a long-term 3D spheroid culture model using MDA-MB-231 cells in a sandwich approach using cell embedding between a non-adherent surface and basement membrane extracts. This allowed consistent growth of spheroids for more than 21 days. Also, co-culturing of MDA-MB-231 with CCD-1137Sk fibroblasts yielded stably growing spheroids, suggesting the importance of extracellular matrix (ECM) in this process. In addition, we have set up a novel and simple open source analysis tool to characterize protein expression in 2D cultures and spheroids by immunofluorescence. Using this approach in combination with Western blot analysis, the expression profile of BSP was analyzed. BSP was enriched at the rims of spheroids, both in mono- and co-cultures and its abundance in general correlated with that of TGFß1 under different conditions, including spheroid maturation, cytostatic treatment, and fibroblast co-culture. Conversely, correlation of IGF-1 and BSP was limited to mono-culture time course profiles. In conclusion, we present novel tools to study the regulation of gene expression in combination with cell proliferation and apoptosis in a long-term 3D model of breast cancer and find dynamic abundance profiles of the metastasis-relevant protein BSP and its regulators.

Carcinoma of unknown primary in the head and neck: comparison between positron emission tomography (PET) and PET/CT.

BACKGROUND: Comparison of the diagnostic validity of positron emission tomography (PET) alone with integrated PET and CT (PET/CT) in the search for occult primary tumors in patients with cancer of unknown primary (CUP) site in the head and neck. METHODS: Thirty-nine consecutive patients with clinical CUP were investigated with PET and 38 patients with PET/CT. After initial diagnostic panendoscopy and histological confirmation of the cervical lymph node metastasis, either PET or PET/CT scanning was performed. RESULTS: Integrated PET/CT had a significantly higher overall detection rate than dedicated PET alone (55.2% vs 30.8%; p = .039) and positive prediction rate (93.3% vs 46.1%; p = .01). CONCLUSION: Integrated PET/CT showed to be superior to PET in the detection of the primary site of clinically occult tumors in CUP syndrome. However, a negative result should still be investigated further by means of panendoscopy with tonsillectomy and blind biopsies.

Primary charge effects on prolate spheroids with moderate aspect ratios.

In this article, we investigate the behavior of charged spheroidal colloids with moderate aspect ratios in linear flow fields. We use direct numerical simulation with body-fitted grids for the solution of the Stokes-Poisson-Nernst-Planck system to include all non-linear effects. Therefore, we propose an efficient semi-implicit time discretization based on a splitting of the Stokes equation. We will study the effects of the electric double layer on the forces, torques and on the motion of spheroidal particles. For low Reynolds numbers, we find approximating linear expressions between the ambient fluid flow and the force and torque on the particle. The description of this linear behavior is based on the resistance functions, whose dependencies on the Debye length and the zeta potential are investigated. It is recovered that the resistance functions obey a quadratic dependence on the zeta potential in the small zeta potential regime. For low values of the zeta potential, approximate formulas for the resistance functions are given. The approximation properties are carefully studied by comparing the approximate results with direct numerical simulations. For the case of a shear flow, the approximate formulas can be used to avoid time-consuming direct numerical simulations.

Investigation of the nonlinear effects during the sedimentation process of a charged colloidal particle by direct numerical simulation.

In this article we study the settling process of a colloidal particle under the influence of a gravitational or centrifugal field in an unbounded electrolyte solution. Since particles in aqueous solutions normally carry a non-zero surface charge, a microscopic electric field develops which alters the sedimentation process compared to an uncharged particle. This process can be mathematically modelled via the Stokes-Poisson-Nernst-Planck system, a system of coupled partial differential equations that have to be solved in an exterior domain. After a dimensional analysis we investigate the influence of the various characteristic dimensionless numbers on the sedimentation velocity. Thereby the linear-response (weak-field) approximation that underpins almost all existing theoretical work on classical electrokinetic phenomena is relaxed, such that no additional assumption on the thickness of the double layer as well as on its displacement is needed. We show that there exists a strong influence of the fluid Reynolds number and the ionic strength on the sedimentation velocity. Further we have developed an asymptotic expansion to describe the limit of small values of the surface potential of a single particle. This expansion incorporates all nonlinear effects and extends the well-known results of Booth (1954) [1] and Ohshima et al. (1984) [2] to higher fluid Reynolds numbers.

Regiodivergent epoxide opening: a concept in stereoselective catalysis beyond classical kinetic resolutions and desymmetrizations.

An approach to highly regiodivergent epoxide openings (REOs) is presented. The very popular kinetic resolutions of epoxides and openings of meso-epoxides constitute subclasses of such REOs. REOs are attractive for parallel resolutions, double asymmetric reactions of enantiomerically enriched epoxides, and for semisynthetic applications in the functionalization of natural products. They have been notoriously difficult to realize by means of SN2 mechanisms. Our titanocene-catalyzed radical REO addresses this issue by decoupling epoxide opening and radical trapping and is firmly based on a mechanistic study of the reductive epoxide opening.

Mechanism of titanocene-mediated epoxide opening through homolytic substitution.

The mechanism of titanocene-mediated epoxide opening was studied by a combination of voltammetric, kinetic, computational, and synthetic methods. With the aid of electrochemical investigations the nature of a number of Ti(III) complexes in solution was established. In particular, the distribution of monomeric and dimeric Ti(III) species was found to be strongly affected by the exact steric conditions. The overall rate constants of the reductive epoxide opening were determined for the first time. These data were employed as the basis for computational studies of the structure and energies of the epoxide-titanocene complexes, the transition states of epoxide opening, and the beta-titanoxy radicals formed. The results obtained provide a structural basis for the understanding of the factors determining the regioselectivity of ring opening and match the experimentally determined values. By employing substituted titanocenes even more selective epoxide openings could be realized. Moreover, by properly adjusting the steric demands of the catalysts and the substrates the first examples of reversible epoxide openings were designed.