The Politics of Race and Class and the Changing Spatial Fortunes of the McCarren Pool in Brooklyn, New York, 1936-2010

This paper explores the changing spatial properties of the McCarren Pool and connects them to the politics of race and class. The pool was a large liberal government project that sought to improve the leisure time of working class Brooklynites and between 1936 and the early 1970s it was a quasi-public functional space. In the 1970s and the early 1980s, the pool became a quasi-public dysfunctional space because the city government reduced its maintenance and staffing levels. Working class whites of the area engaged into neighborhood defense in order to prevent the influx of Latinos and African Americans into parts of Williamsburg and Greenpoint and this included the environs of the McCarren Pool. The pool was shut down in 1983 because of a mechanical failure. Its restoration did not take place because residents and storekeepers near the vicinity of the pool complained that by the 1970s, it was only African Americans and Latinos who patronized the pool and that their presence in the neighborhood undermined white exclusivity. For two decades, the McCarren Pool became a multi-use alternative space frequented by homeless people, graffiti artists, heroin users, teenagers, and drug dealers. Unlike previous decades, during this period, people of various racial and ethnic backgrounds frequented the pool area in a relatively harmonious manner. In the early part of the twenty-first century, a neoliberal city administration allowed a corporation to organize music concerts in the pool premises and promised to restore the facility into an operable swimming pool. The problem with this restoration project is that the history of the pool between the early 1970s and the early 2000s is downplayed and this does not serve well former or future users of the poo

Abstracts from the 20th International Symposium on Signal Transduction at the Blood-Brain Barriers

https://deepblue.lib.umich.edu/bitstream/2027.42/138963/1/12987_2017_Article_71.pd

Nanostructure of DNA repair foci revealed by superresolution microscopy

Induction of DNA double‐strand breaks (DSBs) by ionizing radiation leads to formation of micrometersized DNA‐repair foci, whose organization on the nanometer‐scale remains unknown because of the diffraction limit (~200 nm) of conventional microscopy. Here, we applied diffraction‐unlimited, direct stochastic optical‐reconstruction microscopy (dSTORM) with a lateral resolution of ~20 nm to analyze the focal nanostructure of the DSB marker histone γH2AX and the DNA‐repair protein kinase (DNA‐PK) in irradiated glioblastoma multiforme cells. Although standard confocal microscopy revealed substantial colocalization of immunostained γH2AX and DNA‐PK, in our dSTORM images, the 2 proteins showed very little (if any) colocalization despite their close spatial proximity. We also found that γH2AX foci consisted of distinct circular subunits (“nanofoci”) with a diameter of ~45 nm, whereas DNA‐PK displayed a diffuse, intrafocal distribution. We conclude that γH2AX nanofoci represent the elementary, structural units of DSB repair foci, that is, individual γH2AX‐containing nucleosomes. dSTORM‐based γH2AX nanofoci counting and distance measurements between nanofoci provided quantitative information on the total amount of chromatin involved in DSB repair as well as on the number and longitudinal distribution of γH2AX‐containing nucleosomes in a chromatin fiber. We thus estimate that a single focus involves between ~0.6 and ~1.1Mbp of chromatin, depending on radiation treatment. Because of their ability to unravel the nanostructure of DSB‐repair foci, dSTORM and related single‐molecule localization nanoscopy methods will likely emerge as powerful tools in biology and medicine to elucidate the effects of DNA damaging agents in cells

Migration pattern, actin cytoskeleton organization and response to PI3K-, mTOR-, and Hsp90-inhibition of glioblastoma cells with different invasive capacities

High invasiveness and resistance to chemo- and radiotherapy of glioblastoma multiforme (GBM) make it the most lethal brain tumor. Therefore, new treatment strategies for preventing migration and invasion of GBM cells are needed. Using two different migration assays, Western blotting, conventional and super-resolution (dSTORM) fluorescence microscopy we examine the effects of the dual PI3K/mTOR-inhibitor PI-103 alone and in combination with the Hsp90 inhibitor NVP-AUY922 and/or irradiation on the migration, expression of marker proteins, focal adhesions and F-actin cytoskeleton in two GBM cell lines (DK-MG and SNB19) markedly differing in their invasive capacity. Both lines were found to be strikingly different in morphology and migration behavior. The less invasive DK-MG cells maintained a polarized morphology and migrated in a directionally persistent manner, whereas the highly invasive SNB19 cells showed a multipolar morphology and migrated randomly. Interestingly, a single dose of 2 Gy accelerated wound closure in both cell lines without affecting their migration measured by single-cell tracking. PI-103 inhibited migration of DK-MG (p53 wt, PTEN wt) but not of SNB19 (p53 mut, PTEN mut) cells probably due to aberrant reactivation of the PI3K pathway in SNB19 cells treated with PI-103. In contrast, NVP-AUY922 exerted strong anti-migratory effects in both cell lines. Inhibition of cell migration was associated with massive morphological changes and reorganization of the actin cytoskeleton. Our results showed a cell line-specific response to PI3K/mTOR inhibition in terms of GBM cell motility. We conclude that anti-migratory agents warrant further preclinical investigation as potential therapeutics for treatment of GBM