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Little data exist regarding the practice of sodium management in acute neurologically injured patients. This study describes the practice variations, thresholds for treatment, and effectiveness of treatment in this population. This retrospective, multicenter, observational study identified 400 ICU patients, from 17 centers, admitted for ≥48 h with subarachnoid hemorrhage (SAH), traumatic brain injury (TBI), intraparenchymal hemorrhage, or intracranial tumors between January 1, 2011 and July 31, 2012. Data collection included demographics, APACHE II, Glascow Coma Score (GCS), serum sodium (Na+), fluid rate and tonicity, use of sodium-altering therapies, intensive care unit (ICU) and hospital length of stay, and modified Rankin score upon discharge. Data were collected for the first 21 days of ICU admission or ICU discharge, whichever came first. Sodium trigger for treatment defined as the Na+ value prior to treatment with response defined as an increase of ≥4 mEq/L at 24 h. Sodium-al...

Identification of epitopes defined by T-cell responses aids to (1) monitor antigen-specific cellular immune responses (2) guide rational vaccine design, and (3) understand the nature of protective or harmful T-cell responses in diseases with defined target antigens. The 6-h intracellular cytokine staining (ICS) assay preferentially identifies effector T cells that are readily detectable in the peripheral circulation. In contrast, the whole blood assay (WBA) allows to gauge expansion of antigen-specific T cells over time (7 days), i.e., T cells with lower frequencies (e.g., memory T cells) defined by proliferation and cytokine production. Any cellular immune profile can be measured in the WBA (using the 7 days cell culture supernatants) or directly in responder T cells after antigenic stimulation (in the ICS) with appropriate cytokine-specific detection systems. The choice of the cytokine test panel depends on the nature of the expected immune response. A broad panel of candidate peptides can be tested for T-cell recognition in the WBA due to its simplicity and the low input of (unprocessed, heparinized) blood.

This study examines the regulation of progesterone receptor (PR) in the inhibition of pituitary luteinizing hormone (LH) secretion. Ovariectomized ewes underwent hypothalamic-pituitary disconnection, were pulsed with gonadotropin-releasing hormone (GnRH) and received 1 of 4 treatments: estradiol alone (E), estradiol priming before progesterone (E+P), E removed and replaced with P (E-P), or no steroids (C). P treatment for 24 h, with E or following E-priming, reduced LH pulse amplitude by 55% (p<0.05). E alone did not affect LH release. E increased pituitary cytosolic P binding capacity fourfold over controls (p<0.01) and P further increased binding to eight times controls (p<0.01). Pituitary PR mRNA increased to 149 and 171% of C in E and E+P groups, respectively (p<0.05), but E removal resulted in PR mRNA levels not different from controls. Pituitary receptors for GnRH were tripled by E alone compared to C (p<0.01), whereas P alone or with E had no effect. These data suggest an E-induced, direct pituitary inhibition of LH secretion by P and that this effect of P is associated with E-enhanced binding of P in the pituitary. Additionally, the direct pituitary effects of P on LH secretion cannot be accounted for by influences on GnRH receptor numbers.

Methods are described here to monitor changes in protein level and subcellular localization during the cell cycle progression in the budding yeast Saccharomyces cerevisiae. Cell synchronization is achieved by an α-factor-mediated block-and-release protocol. Cells are collected at different time points for the first two cell cycles upon release. Cellular DNA contents are analyzed by flow cytometry. Trichloroacetic acid protein precipitates are prepared for monitoring levels of cell cycle regulated proteins by Western blotting. The dynamic changes in protein subcellular localization patterns are examined by indirect immunofluorescence microscopy.

1. Introduction Transepithelial chloride secretion requires the activation of both apical mem-brane chloride channels and basolateral membrane potassium channels. Chlo-ride channels are needed for the apical membrane exit of chloride and potassium channels are needed to ...

PCR-based DNA array technology is one of the most suitable techniques to detect and identify multiple pathogens in a single assay. Out of the different array platforms that currently exist, membrane-based DNA macroarrays are the most convenient for plant disease diagnosis because of low costs, great sensitivity, and modest equipment requirements. Here we describe a protocol for routine detection of plant pathogens using DNA macroarrays, i.e., from sampling to analysis of hybridization results. Diagnosis can be completed within 36 h after sample collection.