The successful use of extra-corporeal membrane oxygenation in the management of a pregnant woman with severe H1N1 2009 influenza complicated by pneumonitis and adult respiratory distress syndrome.

We report a case of H1N1 2009 influenza A, in a previously fit woman at 24 weeks of gestation, who presented atypically with abdominal pain. The infection was complicated by severe respiratory failure and acute respiratory distress syndrome, requiring ventilatory support, including extra-corporeal membrane oxygenation (ECMO). This was one of the first cases of severe H1N1 disease presenting in the UK. Use of extra-corporeal membrane oxygenation for the complications of H1N1 resulted in full maternal recovery and subsequent delivery of a healthy infant.

Selection of transfected mammalian cells.

Analysis of gene function frequently requires the formation of mammalian cell lines that contain the studied gene in a stably integrated form. Approximately one in 10(4) cells in a transfection will stably integrate DNA (the efficiency can vary depending on the cell type). Therefore, a dominant, selectable marker is used to permit isolation of stable transfectants. In the first part of this unit, the procedure for determining selection conditions and the resulting stable transfection is presented and the most commonly used selectable markers are discussed. The second protocol includes conditions for thirteen markers commonly used for selection of mammalian cells. A third protocols describes selection of transfected cells from the total population soon after transfection with plasmids that express both the gene of interest and a selection tag. Optimization of transfection conditions can be facilitated by a simple staining assay detailed in a support protocol.

Inhibition of histamine, leukotriene C4/D4, and thromboxane B2 release from human leukocytes and human chopped lung mast cells by the allergic mediator release inhibitor, CI-949.

The novel antiallergy compound, 5-methoxy-3-(1-methylethoxy)-1-phenyl-N-1H-tetrazol-5-yl-1H- indole-2- carboxamide, L-arginine salt (CI-949), inhibited mediator release from human basophilic leukocytes and from human chopped lung mast cells challenged with anti-IgE. In leukocytes, CI-949 was a more potent inhibitor of leukotriene C4/D4 and thromboxane B2 release (concentration of drug that inhibits mediator release by 50% [IC50] 0.5 and 0.1 mumol/L, respectively) than of histamine (IC50, 11.4 mumol/L) when anti-IgE was the challenging stimulus. In human lung, inhibition of release of all three mediators occurred at approximately equal concentrations (IC50s for histamine, 16.6 mumol/L; for leukotriene C4/D4, 7.1 mumol/L; and for thromboxane B2, 6.2 mumol/L). The inhibition of histamine release from basophils by CI-949 was further characterized using a variety of stimuli. Challenge with anti-IgE, histamine-releasing factor derived from lymphocytes, N-formyl-L-methionyl-L-leucyl-L-phenylalanine, and concanavalin A revealed potent inhibition (IC50, 10 to 15 mumol/L). CI-949 was less potent versus calcium ionophore A23187, phorbol myristate acetate (12-o-tetradecanoylphorbol-13-acetate), and C5a (IC50s, 30, 54, and 60 mumol/L, respectively). These results suggest that diverse pathways of cell activation-excitation coupling exist for different stimuli in basophils. Furthermore, the activity and potency of CI-949 in inhibiting release of histamine, leukotrienes, and thromboxane from both human basophils and mast cells suggest that the compound will be effective clinically for indications in which these mediators are implicated, including asthma and allergic rhinitis.

CI-949: a new, potential antiallergy compound inhibits antigen-induced allergic reactions in guinea-pigs in vitro and in vivo.

CI-949 (5-methoxy-3-(1-methyl-ethoxy)-1-phenyl-N-1H-tetrazol-5-yl-1H -indole-2-carboxamide) effectively inhibited the release of histamine and the synthesis or release of immunoreactive sulfidopeptide leukotrienes C4-D4 and thromboxane B2 from antigen-challenged lung fragments of of actively sensitized guinea-pigs. The IC50s were 26.7 +/- 2.8 microM for histamine, 2.7 +/- 2.4 microM for leukotriene, and 3.0 +/- 1.8 microM for thromboxane. Drugs including ketotifen, cromolyn and nedocromil did not inhibit histamine release or did so only at high concentrations, and only cromolyn inhibited the synthesis or releases of the 2 eicosanoids. A dose of 50 mg/kg i.p. of CI-949 protected conscious, aerosol-allergen challenged guinea-pigs for at least 1 h and 100 mg/kg i.p. or per os protected for at least 2 h. These results, the comparisons to standard antiallergic drugs, and other data from experiments with human lung fragments and isolated peripheral leukocytes 1,2,3,4 suggest that CI-949 should be evaluated for clinical activity against allergic and inflammatory conditions in which histamine, sulfidopeptide leukotrienes, and/or thromboxane mediate symptoms.

Novel indolecarboxamidotetrazoles as potential antiallergy agents.

The synthesis and antiallergic potential of a series of novel indolecarboxamidotetrazoles are described. A number of compounds inhibit the release of histamine from anti-IgE-stimulated basophilic leukocytes obtained from allergic donors. Optimal inhibition is exhibited by compounds with 3-alkoxy, 5-methoxy, and 1-phenyl substituents on the indole core structure. Compound 8d (5-methoxy-3-(1-methylethoxy)-1-phenyl-N-1H-tetrazol-5-yl-1H -indole-2-carboxamide; designated CI-949) is a potent inhibitor of histamine release from human basophils and from guinea pig and human chopped lung.

Clinical evaluation of intravenous nitroglycerin for neurosurgery.

Moment-to-moment control of blood pressure is important in the management of the neurosurgical patient. The ideal agent to control blood pressure or induce hypotension should be non-toxic, maintain cerebrovascular autoregulation, and not alter cardiac output or change intracranial pressure. Intravenous nitroglycerin has been used to control blood pressure in 54 neurosurgical cases. This agent produces a rapid, controllable, but not precipitous fall in blood pressure without rebound, is non-toxic, may not alter cerebrovascular autoregulation, and does not raise intracranial pressure. Our clinical experience with intravenous nitroglycerin indicates that it has an important role as a hypotensive agent for the neurosurgical patient.

Air embolism during trans-sphenoidal pituitary operations.

Doppler ultrasonic cardiac monitoring of patients undergoing trans-sphenoidal pituitary operations in the semisitting position has revealed the occurrence of venous air embolism in 3 of 31 consecutive cases. One such case is presented. Air may be drawn into the venous system whenever a gradient exists between the site of operation and the right heart. During trans-sphenoidal operations the most likely portals of venous air entry include the intercavernous connections within the sella, venous channels through nonpneumatized bone, inadequately sealed subnasal vessels, and vascularized metastatic tissue in the pituitary. Because the potential for morbidity and mortality from air embolism is so great, rapid diagnosis with the Doppler unit and prompt treatment, including aspiration of air from the right atrial catheter, administration of 100% oxygen, performance of the Valsalva maneuver, saline irrigation of the wound, and hemostasis of open vessels, are essential. Technetium-macroaggregated albumin (TEMAA) lung scans are helpful in postoperative verification of venous air embolism.