Topoisomerase II (topo II) inhibitors are commonly used as chemotherapy to treat multiple types of cancer, though their use is also associated with the development of therapy related acute leukemias. While the chromosome-damaging effects of etoposide, a topo II poison, have been proposed to act through a threshold mechanism, little is known about the chromosome damaging effects and dose responses for the catalytic inhibitors of the enzyme. The current study was designed to further investigate the potencies and concentration-response relationships of several topoisomerase II inhibitors, including the topoisomerase II poison etoposide, as well as catalytic inhibitors aclarubicin, merbarone, ICRF-154 and ICRF-187 using both a traditional in vitro micronucleus assay as well as a flow-cytometry based version of the assay. Benchmark dose (BMD) analysis was used to identify models that best fit the data and estimate a BMD, in this case the concentration at which a one standard deviation increase above the control frequency would be expected. All of the agents tested were potent in inducing micronuclei in human lymphoblastoid TK6 cells, with significant increases seen at low micromolar, and in the cases of aclarubicin and etoposide, at low nanomolar concentrations. Use of the anti-kinetochore CREST antibody with the microscopy-based assay demonstrated that the vast majority of the micronuclei originated from chromosome breakage. In comparing the two versions of the micronucleus assay, significant increases in micronucleated cells were observed at similar or lower concentrations using the traditional microscopy-based assay. BMD modeling of the data exhibited several advantages and proved to be a valuable alternative for concentration-response analysis, producing points of departure comparable to those derived using traditional no-observed or lowest-observed genotoxic effect level (NOGEL or LOGEL) approaches.

We report direct evidence that magnetically charged superdomain walls form spontaneously in two dimensional square artificial spin ice nanostructures in response to external magnetic fields. These extended magnetic defects were revealed by the development of internal structure, which varies as a function of applied magnetic field, within the Bragg peaks of resonant soft x-ray magnetic scattering patterns. Magnetically charged superdomain walls extend over tens of lattice sites and do not necessarily align with the applied field. Our results illustrate a novel approach to detect hierarchical magnetic structures within spin textures.

We report evidence of spontaneous formation of a heterogeneous network of superdomains in two-dimensional square artificial spin ice nanostructures in externally applied magnetic fields. Coherent resonant soft-x-ray scattering from such textures gives rise to unique internal structure in Bragg peaks. The magnetic heterogeneity is locally disordered but has a zigzag texture at longer length scales. Our result suggests that the macroscopic magnetic texture is derived from the microscopic structure of the Dirac strings.