A marker-independent lineage-tracing system to quantify clonal dynamics and stem cell functionality in cancer tissue

Lineage tracing is a powerful tool that can be used to uncover cell fates. Here, we describe a novel method for the quantitative analysis of clonal dynamics in grafted cancer tissues. The protocol involves the preparation and validation of cells for lineage tracing, establishment of grafts and label induction, analysis of clone-size distribution and fitting of the experimental data to a mathematical tumor growth model. In contrast to other lineage-tracing strategies, the method described here assesses stem cell functionality and infers tumor expansion dynamics independently of molecular markers such as putative cancer stem cell (CSC)-specific genes. The experimental system and analytical framework presented can be used to quantify clonal advantages that specific mutations provide, in both the absence and presence of (targeted) therapeutic agents. This protocol typically takes ~20 weeks to complete from cell line selection to inference of growth dynamics, depending on the grafted cancer growth rate

The role of iron in the formation of porosity in Al-Si-Cu based alloys - Part I: Initial experimental observations

Outbreaks of interconnected porosity in industrial Al-Si-based alloy castings have, on occasion, been attributed to variations in metal chemistry rather than to changes in process parameters. This work identifies the role that iron plays in porosity formation and reports a threefold effect in an Al-5 pct Si-1 pct Cu-0.5 pct Mg alloy. In addition to a detailed analysis of casting porosity profiles, metallographic and thermal studies also point to inadequacies in the existing theories regarding the role of iron and suggest that a new theory is required to understand the observed behavior