Localization of acetylcholine receptors and cholinesterase on nerve-contacted and noncontacted muscle cells grown in the presence of agents that block action potentials

The possible role of nerve activity in triggering changes in the localization of acetylcholine receptors (AChRs) and cholinesterase (ChE) on nerve-contacted Xenopus muscle cells has been assessed. The localization of these molecules was examined on nerve-contacted and noncontacted muscle cells in cultures of spinal cord and myotomal muscle derived from Xenopus embryos. Sites of high AChR density were revealed by staining with fluorescent alpha-bungarotoxin and sites of ChE localization were revealed histochemically. Localization of AChRs and ChE at sites of nerve-muscle contact occurred when the culture medium contained 1.2 micron tetrodotoxin (TTX), 1.2 micron TTX, 10 mM magnesium, and no calcium salts, 1.2 micron TTX and 2 mM manganese, or 106 mM potassium methyl sulfate instead of sodium chloride. The nerve-contacted muscle cells in each of these modified culture media also exhibited a reduced incidence of AChR and ChE patches away from the site of contact. It is concluded that the neural factor(s) that triggers the local and remote changes in AChR and ChE distribution can be supplied to the neurites and externalized in the absence of nerve impulses, and that the nerve and muscle cells can interact even when they are largely depolarized.     

Methodological issues in drug-drug conditioning in rats: nonassociative factors in heart rate and avfail.

Sodium pentobarbital injections followed 30 min later by d-amphetamine sulfate produce an effect over trials in the form of an increase in heart rate in response to pentobarbital in relation to rats that receive the 2 drugs 24 hr apart (long-delayed control: Revusky, Davey, & Zagorski, 1989). This study found equivalent increases in heart rate in forward and backward groups in relation to a long-delayed control regardless of whether training or testing was carried out in a heart rate recording apparatus or in the home cage, which suggests that a drug interaction due to drug administrations in forward and backward groups has yet to be eliminated in accounting for the heart rate effect. Comparison of backward and long-delayed controls in a drug-drug procedure that used a taste aversion test revealed that both forward and delayed pairings can produce attenuated aversions in relation to a backward group regardless of whether the unconditional stimulus is amphetamine (Experiment 1) or lithium chloride (Experiment 2).     

Emergency use of the laryngeal mask airway in severe upper airway obstruction caused by supraglottic oedema

We report two cases of severe upper airway obstruction caused by supraglottic oedema which developed rapidly at the time of anaesthesia. Conventional methods to relieve the obstruction failed and it was only overcome when a laryngeal mask airway (LMA) was inserted and positive pressure applied manually during inspiration. In one case a fibrescope was passed via the LMA and this revealed two cushions of oedematous false vocal cords protruding into the bowel of the LMA which were pushed out of the way when positive pressure was applied during inspiration. We believe that the LMA should be considered in the emergency management of severe upper airway obstruction even when this involves supraglottic oedema.      

Tolerance of Pseudomonas aeruginosa to killing by ciprofloxacin, gentamicin and imipenem in vitro and in vivo

In the rat croton oil pouch model, treatment with ciprofloxacin, gentamicin or imipenem caused a sharp reduction in the numbers of Pseudomonas aeruginosa when applied 24 h after infection but treatment had little or no effect when delayed until 48 h after infection. Surviving bacteria from treated animals were fully susceptible on subculture in vitro. Between 24 h and 48 h after infection there was an increase in the concentration of magnesium and a decrease in the concentration of oxygen but no significant change in the pH of the pouch fluid. In-vitro, the MBC of gentamicin was increased four-fold by the addition of magnesium ions and eight-fold under anaerobic conditions whereas these variables had no significant effect upon the MBC of ciprofloxacin and imipenem. However, cells of P. aeruginosa incubated in batch culture for 4, 24 and 48 h became progressively more tolerant to killing by all three drugs. We conclude that the tolerance of P. aeruginosa in vivo was only partly explained by biochemical changes at the site of infection. The stationary growth phase was associated with progressive tolerance to killing by ciprofloxacin, gentamicin and imipenem both in vivo and in vitro.     

Preselection Thymocytes Are More Sensitive to T Cell Receptor Stimulation Than Mature T Cells

During T cell development, thymocytes which are tolerant to self-peptides but reactive to foreign peptides are selected. The current model for thymocyte selection proposes that self-peptide–major histocompatibility complex (MHC) complexes that bind the T cell receptor with low affinity will promote positive selection while those with high affinity will result in negative selection. Upon thymocyte maturation, such low affinity self-peptide–MHC ligands no longer provoke a response, but foreign peptides can incidentally be high affinity ligands and can therefore stimulate T cells. For this model to work, thymocytes must be more sensitive to ligand than mature T cells. Contrary to this expectation, several groups have shown that thymocytes are less responsive than mature T cells to anti-T cell receptor for antigen (TCR)/CD3 mAb stimulation. Additionally, the lower TCR levels on thymocytes, compared with T cells, would potentially correlate with decreased thymocyte sensitivity. Here we compared preselection thymocytes and mature T cells for early activation events in response to peptide–MHC ligands. Remarkably, the preselection thymocytes were more responsive than mature T cells when stimulated with low affinity peptide variants, while both populations responded equally well to the antigenic peptide. This directly demonstrates the increased sensitivity of thymocytes compared with T cells for TCR engagement by peptide–MHC complexes.