Replication Labeling Methods for Super-Resolution Imaging of Chromosome Territories and Chromatin Domains.

Continuing progress in super-resolution microscopy enables the study of sub-chromosomal chromatin organization in single cells with unprecedented detail. Here we describe refined methods for pulse-chase replication labeling of individual chromosome territories (CTs) and replication domain units in mammalian cell nuclei, with specific focus on their application to three-dimensional structured illumination microscopy (3D-SIM). We provide detailed protocols for highly efficient electroporation-based delivery or scratch loading of cell-impermeable fluorescent nucleotides for live-cell studies.

Chromatin arranges in chains of mesoscale domains with nanoscale functional topography independent of cohesin.

Three-dimensional (3D) chromatin organization plays a key role in regulating mammalian genome function; however, many of its physical features at the single-cell level remain underexplored. Here, we use live- and fixed-cell 3D super-resolution and scanning electron microscopy to analyze structural and functional nuclear organization in somatic cells. We identify chains of interlinked ~200- to 300-nm-wide chromatin domains (CDs) composed of aggregated nucleosomes that can overlap with individual topologically associating domains and are distinct from a surrounding RNA-populated interchromatin compartment.

Anthropometrical and Physical Fitness Predictors of Operational Military Test Performance in Air Force Personnel.

The objective of this study was to verify the relationships between the anthropometrical and physical fitness parameters (measured by the Physical Conditioning Assessment (PCA) of the Aeronautics Command), with the operational performance in the simulated military task performance (SMTP) performed by the infantry military of a Brazilian Air Force (BAF) unit. These evaluations were performed on two distinct days, interspersed by 48h, with PCA on the first day and the SMTP in the second. The distribution of the dependent variable was not normal (Shapiro-Wilk test, = 0.

Stabilization of chromatin topology safeguards genome integrity.

To safeguard genome integrity in response to DNA double-strand breaks (DSBs), mammalian cells mobilize the neighbouring chromatin to shield DNA ends against excessive resection that could undermine repair fidelity and cause damage to healthy chromosomes. This form of genome surveillance is orchestrated by 53BP1, whose accumulation at DSBs triggers sequential recruitment of RIF1 and the shieldin-CST-POLα complex. How this pathway reflects and influences the three-dimensional nuclear architecture is not known.

[NOTES FROM A SIX MONTHS VOLUNTEER MISSION AS MEDICAL STAFF IN A DISTRICT HOSPITAL IN UGANDA].

In December 2014 my husband and I flew for a stay of six months in Uganda. We went there in order to volunteer in the district hospital of Kiboga, one of the most impoverished districts of this poverty stricken country. A district in which over 60% of the population lives off less than a dollar and twenty five cents a day, in which the average life expectancy is 46.