Density dependent regulation of inflammatory responses in macrophages

Macrophage distribution density is tightly regulated within the body, yet the importance of macrophage crowding during in vitro culture is largely unstudied. Using a human induced pluripotent stem cell (iPSC)-derived macrophage model of tissue resident macrophages, we characterize how increasing macrophage culture density changes their morphology and phenotype before and after inflammatory stimulation. In particular, density drives changes in macrophage inflammatory cytokine and chemokine secretion in both resting and activated states. This density regulated inflammatory state is also evident in blood monocyte derived-macrophages, the human monocytic THP-1 immortalized cell line, and iPSC-derived microglia. Density-dependent changes appear to be driven by a transferable soluble factor, yet the precise mechanism remains unknown. Our findings highlight cell plating density as an important but frequently overlooked consideration of in vitro macrophage research relevant to a variety of fields ranging from basic macrophage cell biology to disease studies

The Use of Digital Terrain Models to Estimate the Pace of Filling the Pit of a Central European Granite Quarry with Water

This paper presents the results of an analysis of the pace of filling one of the deepest European granite quarries with water. A DTM (digital terrain model) based on data from LiDAR ALS (light detection and ranging airborne laser scanning) was used to create a model of the pit of the Strzelin I granite quarry and to determine the reach and surface area of the direct catchment of the excavation pit. The increase in the volume of water in the excavation pit was determined. Analogue maps and DTM were used to calculate the maximum depth of the pit (113.3 m), its surface area (9.71 ha), and its capacity (5.1 million m3). The volume of water collected in the excavation pit during the years 2011–2018 was determined based on the analogue base map and the DTM. The result was 0.335 million m3. Based on the data made available by the mining company, the correlation of the DTM with the orthophotomap of the mining area and additional field measurements, the ordinates of the water level in the years 2011–2018 were determined. Initially, the water surface level in the quarry was located on the ordinate of 66.6 m a.s.l. (July 20, 2011). After the pumping of water was discontinued, the level rose to 96.1 m a.s.l. (January 28, 2018). The increase in the water volume in the quarry pit during specific periods was determined (actual retention increase). The obtained data on the volume of the retained water referred to the period during which it accumulated in the quarry. On average, the net increase in water retention in the excavation pit was 138.537 m3∙d−1, and the calculated net supply from the direct catchment (16.04 ha) was 101.758 m3∙d−1. The use of DTM and measurements of the water level in the excavation pit seem to be an efficient means of estimating the pace of spontaneous filling of the quarry with water supplied from the direct physiographic catchment

Controlling distinct signaling states in cultured cancer cells provides a new platform for drug discovery

Cancer cells can switch between signaling pathways to regulate growth under different conditions. In the tumor microenvironment, this likely helps them evade therapies that target specific pathways. We must identify all possible states and utilize them in drug screening programs. One such state is characterized by expression of the transcription factor Hairy and Enhancer of Split 3 () and sensitivity to knockdown, and it can be modeled . Here, we cultured 3 primary human brain cancer cell lines under 3 different culture conditions that maintain low, medium, and high expression and characterized gene regulation and mechanical phenotype in these states. We assessed gene expression regulation following knockdown in the -high conditions. We then employed a commonly used human brain tumor cell line to screen Food and Drug Administration (FDA)-approved compounds that specifically target the -high state. We report that cells from multiple patients behave similarly when placed under distinct culture conditions. We identified 37 FDA-approved compounds that specifically kill cancer cells in the high--expression conditions. Our work reveals a novel signaling state in cancer, biomarkers, a strategy to identify treatments against it, and a set of putative drugs for potential repurposing.-Poser, S. W., Otto, O., Arps-Forker, C., Ge, Y., Herbig, M., Andree, C., Gruetzmann, K., Adasme, M. F., Stodolak, S., Nikolakopoulou, P., Park, D. M., Mcintyre, A., Lesche, M., Dahl, A., Lennig, P., Bornstein, S. R., Schroeck, E., Klink, B., Leker, R. R., Bickle, M., Chrousos, G. P., Schroeder, M., Cannistraci, C. V., Guck, J., Androutsellis-Theotokis, A. Controlling distinct signaling states in cultured cancer cells provides a new platform for drug discovery