Tetraplegia is associated with increased hypoxic ventilatory response during nonrapid eye movement sleep

Abstract People with cervical spinal cord injury (SCI) are likely to experience chronic intermittent hypoxia while sleeping. The physiological effects of intermittent hypoxia on the respiratory system during spontaneous sleep in individuals with chronic cervical SCI are unknown. We hypothesized that individuals with cervical SCI would demonstrate higher short‐ and long‐term ventilatory responses to acute intermittent hypoxia (AIH) exposure than individuals with thoracic SCI during sleep. Twenty participants (10 with cervical SCI [9 male] and 10 with thoracic SCI [6 male]) underwent an AIH and sham protocol during sleep. During the AIH protocol, each participant experienced 15 episodes of isocapnic hypoxia using mixed gases of 100% nitrogen (N2) and 40% carbon dioxide (CO2) to achieve an oxygen saturation of less than 90%. This was followed by two breaths of 100% oxygen (O2). Measurements were collected before, during, and 40 min after the AIH protocol to obtain ventilatory data. During the sham protocol, participants breathed room air for the same amount of time that elapsed during the AIH protocol and at approximately the same time of night. Hypoxic ventilatory response (HVR) during the AIH protocol was significantly higher in participants with cervical SCI than those with thoracic SCI. There was no significant difference in minute ventilation (V.E.), tidal volume (V.T.), or respiratory frequency (f) during the recovery period after AIH in cervical SCI compared to thoracic SCI groups. Individuals with cervical SCI demonstrated a significant short‐term increase in HVR compared to thoracic SCI. However, there was no evidence of ventilatory long‐term facilitation following AIH in either group

Activation-Induced Deaminase Cloning, Localization, and Protein Extraction from Young VH-Mutant Rabbit Appendix

Studies in mouse, human, and chicken suggest that activation-induced deaminase (AID) is involved in three known processes leading to antibody diversification: somatic hypermutation, gene conversion, and class-switch recombination. Developing rabbit appendix provides a particularly good site for studying all three of these B cell maturation events. We report here successful cloning of rabbit AID and isolation of AID protein from rabbit appendix-cell nuclear and cytoplasmic extracts. We succeeded in identifying and locating AID protein in cells by immunohistochemical and immunofluorescent staining techniques and examined colocalization of AID and other molecules important for Ab diversification. This report extends our knowledge about AID to a mammalian species that uses gene conversion to diversify rearranged Ig genes. Although much work remains to understand fully the mechanism of action of AID and its association with other cellular components, the rabbit system now offers a particularly useful model for future studies of these dynamics

Effect of arginine mutation on EBA-175 RII binding to human erythrocytes.

a<p>the rosette number was an average counted from two biological repeats in each experiment.</p>b<p>erythrocyte-binding assay was performed with normal erythrocytes in two independent experiments.</p>c<p>erythrocyte-binding assay was performed with neuraminidase-treated erythrocytes Nm, neuraminidase.</p>*<p>Residue 566 of EBA-175 corresponds to residue 422 of rEBA-175 RII.</p

Mapping of epitope predictions to the rEBA-175 RII crystal structure.

<p>(A) rEBA-175 RII crystal structure as a dimer of two RII molecules (cyan and grey), the region corresponding to the F2βf peptide is highlighted in yellow and R422 is highlighted in magenta. The amino- and carboxy- terminal residues are colored in blue and red respectively. The white box represents the region highlighted in panel B. (B) The F2 domain of one monomer in the rEBA-175 RII crystal structure is shown as a ribbon diagram and residues are colored by the number of times they were predicted by either PepSurf and Mapitope to be part of the epitope for mAbs R215, R217, or R256 from never (cyan) to most often (magenta; see SOM for raw data). Residues discussed in the text are labeled according to rEBA-175 RII numbering (EBA-175 3D7 sequence number –144) and shown in stick representation.</p

Summary of binding properties of EBA-175 RII specific mAbs.

<p>SPR: surface plasmon resonance. ND: not determined.</p>*<p>Does not inhibit parasite growth <i>in vitro</i><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0056326#pone.0056326-Sim2" target="_blank">[21]</a>.</p>**<p>None of the mAbs demonstrated binding to linear F2βf.</p>c<p>These mAbs were shown to compete against each other for binding by competition ELISA.</p

F2βf peptide competes with mAbs R215 (A) and R217 (B) but not R218 (C) for binding to recombinant EBA-175 RII by Surface Plasmon Resonance studies.

<p>Monoclonal antibody sensograms show binding to immobilized EBA-175 RII with the concentrations of mAb and cyclic F2βf peptide as indicated. The concentrations of cyclic F2βf peptide were added to a fixed concentration of mAb.</p

Deduced amino acid sequences of clones enriched for each EBA-175 RII specific mAb.

<p>Deduced amino acid sequences of clones enriched for each EBA-175 RII specific mAb.</p

The Epitope of Monoclonal Antibodies Blocking Erythrocyte Invasion by <em>Plasmodium falciparum</em> Map to The Dimerization and Receptor Glycan Binding Sites of EBA-175

<div><p>The malaria parasite, <em>Plasmodium falciparum</em>, and related parasites use a variety of proteins with Duffy-Binding Like (DBL) domains to bind glycoproteins on the surface of host cells. Among these proteins, the 175 kDa erythrocyte binding antigen, EBA-175, specifically binds to glycophorin A on the surface of human erythrocytes during the process of merozoite invasion. The domain responsible for glycophorin A binding was identified as region II (RII) which contains two DBL domains, F1 and F2. The crystal structure of this region revealed a dimer that is presumed to represent the glycophorin A binding conformation as sialic acid binding sites and large cavities are observed at the dimer interface. The dimer interface is largely composed of two loops from within each monomer, identified as the F1 and F2 β-fingers that contact depressions in the opposing monomers in a similar manner. Previous studies have identified a panel of five monoclonal antibodies (mAbs) termed R215 to R218 and R256 that bind to RII and inhibit invasion of erythrocytes to varying extents. In this study, we predict the F2 β-finger region as the conformational epitope for mAbs, R215, R217, and R256, and confirm binding for the most effective blocking mAb R217 and R215 to a synthetic peptide mimic of the F2 β-finger. Localization of the epitope to the dimerization and glycan binding sites of EBA-175 RII and site-directed mutagenesis within the predicted epitope are consistent with R215 and R217 blocking erythrocyte invasion by <em>Plasmodium falciparum</em> by preventing formation of the EBA-175– glycophorin A complex.</p> </div