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.

Strategic and practical guidelines for successful structured illumination microscopy.

Linear 2D- or 3D-structured illumination microscopy (SIM or3D-SIM, respectively) enables multicolor volumetric imaging of fixed and live specimens with subdiffraction resolution in all spatial dimensions. However, the reliance of SIM on algorithmic post-processing renders it particularly sensitive to artifacts that may reduce resolution, compromise data and its interpretations, and drain resources in terms of money and time spent. Here we present a protocol that allows users to generate high-quality SIM data while accounting and correcting for common artifacts.

Quantitative 3D structured illumination microscopy of nuclear structures.

3D structured illumination microscopy (3D-SIM) is the super-resolution technique of choice for multicolor volumetric imaging. Here we provide a validated sample preparation protocol for labeling nuclei of cultured mammalian cells, image acquisition and registration practices, and downstream image analysis of nuclear structures and epigenetic marks. Using immunostaining and replication labeling combined with image segmentation, centroid mapping and nearest-neighbor analyses in open-source environments, 3D maps of nuclear structures are analyzed in individual cells and normalized to fluorescence standards on the nanometer scale.

In Vivo and In Situ Replication Labeling Methods for Super-resolution Structured Illumination Microscopy of Chromosome Territories and Chromatin Domains.

Recent advances in super-resolution microscopy enable the study of subchromosomal 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.

Growth parameters of discordant preterm twins during the first year of life.

Objective: To evaluate and compare growth parameters (weight, length and head circumference) of discordant preterm twins during the first year of life.

Methods: Retrospective data were collected on 78 pairs of >10% discordant preterm twins. Data regarding short-term neonatal outcome were recorded.

From service provision to function based performance - perspectives on public health systems from the USA and Israel.

If public health agencies are to fulfill their overall mission, they need to have defined measurable targets and should structure services to reach these targets, rather than offer a combination of ill-targeted programs. In order to do this, it is essential that there be a clear definition of what public health should do- a definition that does not ebb and flow based upon the prevailing political winds, but rather is based upon professional standards and measurements.The establishment of the Essential Public Health Services framework in the U.

Factors affecting Varicella vaccine uptake among children 1-17 years old in the Haifa District in 2007.

The Israeli National Vaccination Program (INVP) is aimed mainly at the pediatric age group. Till 2008 the program did not include routine administration of vaccine against Varicella (chickenpox). For the past 7 years, however, vaccine against Varicella has been available to the public and subsidized through the health maintenance organizations (HMOs).

Describing the social environment of elderly persons at the time of death.

Objectives: To describe the social environment of elderly persons at the time of their death and its correlates.

Design: Telephone interviews with the next-of-kin of deceased elderly persons.

Participants: Potential participants were the next-of-kin of all Jewish elderly individuals who died during the course of one year in a major city in the northern part of Israel.

Push-pull optical pumping of pure superposition states.

A new optical pumping method, "push-pull pumping," can produce very nearly pure, coherent superposition states between the initial and the final sublevels of the important field-independent 0-0 clock resonance of alkali-metal atoms. The key requirement for push-pull pumping is the use of D1 resonant light which alternates between left and right circular polarization at the Bohr frequency of the state. The new pumping method works for a wide range of conditions, including atomic beams with almost no collisions, and atoms in buffer gases with pressures of many atmospheres.

Oral anticoagulants in the elderly.

Over a 1-year period, 34,998 prothrombin time tests were performed on 2,379 patients aged 40-89 years with a recommended target International Normalized Ratio (INR) value of 2.5. At least one INR value of >/= 5 was found in 507/2,379 patients (21.

Population colorectal cancer screening with fecal occult blood test.

Screening for early detection of colorectal cancer using fecal occult blood testing has been shown to be effective in reducing mortality from this disease. The largest Health Maintenance Organization in Israel initiated the use of Hemoccult Sensa in 1992 to evaluate the field performance of this test. All primary care physicians were invited to order home-based tests for their asymptomatic patients 50-74 years of age.

[Shy-Drager syndrome].

An 81-year-old man with a history of chronic pulmonary disease due to heavy smoking and ischemic heart disease had been suffering for the past few years from chronic constipation and urinary incontinence and was receiving medication for cardiopulmonary symptoms and urinary incontinence. He was admitted for repeated falling for a few months prior to admission. When put in the supine position, his blood pressure fell.

Characteristics of 1, 1',3,3,3',3'-hexamethylindotricarbocyanine iodide as a tunable dye laser in the near infrared.

We describe the lasing characteristics of a 1,1',3,3,3',3'-hexamethylindotricarbocyanine iodide infrared dye laser. The absorption spectrum of the dye dissolved in ethanol has a large peak centered at 744 nm. The fluorescence exhibits a Stokes shift of approximately 36 nm with a lifetime of 1.

Self-calibrating technique for beam divergence measurements of cw and pulsed lasers.

A convenient method to measure beam divergence of laser beams is described. The apparatus is compact and accurate due to the use of a telescope to focus the beam. Incorporation of a Fabry-Perot etalon permits focusing and in situ calibration.