Dendritic and synaptic effects in systems of coupled cortical oscillators

We explore the influence of synaptic location and form on the behavior of networks of coupled cortical oscillators. First, we develop a model of two coupled somatic oscillators that includes passive dendritic cables. Using a phase model approach, we show that the synchronous solution can change from a stable solution to an unstable one as the cable lengthens and the synaptic position moves further from the soma. We confirm this prediction using a system of coupled compartmental models. We also demonstrate that when the synchronous solution becomes unstable, a bifurcation occurs and a pair of asynchronous stable solutions appear, causing a phase lag between the cells in the system. Then using a variety of coupling functions and different synaptic positions, we show that distal connections and broad synaptic time courses encourage phase lags that can be reduced, eliminated, or enhanced by the presence of active currents in the dendrite. This mechanism may appear in neural systems where proximal connections could be used to encourage synchrony, and distal connections and broad synaptic time courses could be used to produce phase lags that can be modulated by active currents.     

Water-soluble nicotinic acetylcholine receptor formed by alpha7 subunit extracellular domains

Water-soluble models of ligand-gated ion channels would be advantageous for structural studies. We investigated the suitability of three versions of the N-terminal extracellular domain (ECD) of the alpha7 subunit of the nicotinic acetylcholine receptor (AChR) family for this purpose by examining their ligand-binding and assembly properties. Two versions included the first transmembrane domain and were solubilized with detergent after expression in Xenopus oocytes. The third was truncated before the first transmembrane domain and was soluble without detergent. For all three, their equilibrium binding affinities for alpha-bungarotoxin, nicotine, and acetylcholine, combined with their velocity sedimentation profiles, were consistent with the formation of native-like AChRs. These characteristics imply that the alpha7 ECD can form a water-soluble AChR that is a model of the ECD of the full-length alpha7 AChR.     

Inhibition of IL-1 induced tissue factor (TF) synthesis and procoagulant activity (PCA) in purified human monocytes by IL-4, IL-10 and IL-13

Exposure of monocytes to pro-inflammatory cytokines or lipopolysaccharide (LPS) may induce synthesis and expression of tissue factor (TF). In this paper we have focused on the induction of TF-activity in human monocytes by the pro-inflammatory cytokines recombinant human interleukin 1 (rhIL-1 alpha) (rhIL-1 beta) (rhIL-6) and human tumour necrosis factor alpha (rhTNF-alpha), measured as procoagulant activity (PCA) in a microtitre plate-based clot assay. In addition we have studied the modulation of IL-1 alpha/beta induced TF-mRNA and PCA by rhIL-4, rhIL-10 and rhIL13. IL-1 alpha and IL-1 beta induced a concentration dependent increase in TF-activity. Neither IL-6 nor TNF-alpha gave rise to procoagulant activity at the concentrations tested (0.2-20 ng/ml). IL-4, IL-10 and IL-13, all effectively diminished IL-1 alpha/beta induced PCA, shown at the protein- and at the mRNA-level, while cell viability was unaffected. These results add to the previously demonstrated role of IL-4 and IL-10 as inhibitors of LPS-induced TF-activity, showing that these anti-inflammatory cytokines are not specific for LPS-activation but interfere with other stimulating substances such as IL-1, which may be involved in diseases where LPS is not present.     

Noxiustoxin 2, a novel K+ channel blocking peptide from the venom of the scorpion Centruroides noxius Hoffmann

A novel peptide called Noxiustoxin 2 (NTX2) was purified from the venom of the scorpion Centruroides noxius and characterized chemically and functionally. It is composed of 38 amino acid residues linked by three disulfide bridges and its primary structure is 61% identical to that of Noxiustoxin (NTX). It is not toxic to mice (using up to 200 micrograms/20 g mouse weight) and crustaceans (up to 30 micrograms/g of crayfish), but has a paralysing effect on crickets (30 micrograms/g animal). It displaces the binding of [125I]NTX to rat brain synaptosome membranes with a Ki of 0.1 microM, in comparison NTX has a Ki of 100 pM. Similarly, using single Ca2+ activated K+ channels of small conductance obtained from cultured bovine aortic endothelial cells it was shown that NTX2 is over two logarithm units less potent than NTX in producing 50% blockade of the probability of opening the channels. NTX2 is not recognized by a panel of six distinct monoclonal antibodies against NTX, however it is recognized by polyclonal antibodies raised in mouse, with native NTX. Primary structure comparison of both NTX and NTX2 suggests that the N-terminal segments of these peptides are important for channel affinity.     

Alteration by phosphatidyl serine of tension responses and 45Ca distribution in aortic smooth muscle

The effects of phosphatidyl serine (PS) on 45Ca distribution, 45Ca movements and contractions were examined in rabbit aortic smooth muscle. Contractile responses to submaximal concentrations of norepinephrine and histamine were potentiated by prior exposure to PS, but equivalent responses to potassium were unaffected. Addition of PS to the incubation solution decreased 45Ca uptake; exposure of aortic strips to PS during washout of either 45Ca or promethium (147Pm) resulted in maintained increases in efflux. These PS-induced alterations in net loss of 45Ca or 147Pm can be attributed to a decreased membrane reuptake and/or rebinding. However, the presence of PS during the washout significantly reduced the increases in 45Ca efflux rate elicited with either 0.05 mM concentrations of Ca++ or ethylenediamine tetraacetic acid. Thus, in rabbit aortic smooth muscle, exogenous PS can alter the availability and/or exchangeability of a membrane-bound Ca++ fraction. By specifically increasing the affinity for Ca++ at relevant membrane sites or stores. PS may enhance the ability of vascular smooth muscle to respond to stimulatory agents that mobilize Ca++ from these sites and, in this manner, potentiate contractile responses.     

In vitro activity of guinea pig transfer factor released into plasma

Plasma fractions and plasma dialysate from 2,4-dinitrochlorobenzene- and tuberculin-sensitive guinea pigs that had been treated with either antilymphocytic serum or normal control serum were analyzed for their ability to transfer lymphocyte transformation, passive cutaneous anaphylaxis, and macrophage migration inhibition, as well as delayed hypersensitivity in vivo. Antilymphocytic serum caused rapid release of material, which has characteristics of transfer factor, into the plasma. It was dialyzable, migrated electrophoretically with the alpha globulins and albumin, possessed a 280/260 (nm) optical density ratio of 0.7, and caused in vitro lymphocyte transformation in the presence of the specific antigen. Passive cutaneous anaphylaxis antibodies were also present in the plasma of sensitive animals, but they were isolated in electrophoretic or dialysis fractions separate from those containing transfer activity.     

Schistosoma mansoni: an enkephalinergic system that may participate in internal and host-parasite signaling

The present study is concerned with an opioid system in the human trematode Schistosoma mansoni, both as part of the endogenous chemical messenger system and as a tool in the parasite reaction to the host(s). A high-affinity opioid binding site was characterized in membrane suspensions prepared from adult worms. Scatchard analysis revealed a single class of receptors with a dissociation constant of 1.8 nM and a Bmax of 24.9 pmol/g protein for (D-Ala2, Met5)-enkephalin (DAME). The displacement experiments demonstrated that the most potent ligands were beta-endorphin, DAME, and met-enkephalin. These characteristics and the effects of various ions on DAME affinity suggest that S. mansoni has a delta-like opioid receptor, as previously described in other invertebrates. A met-enkephalin-like peptide was also characterized in a miracidial extract. Radioimmunoassay, reverse-phase HPLC, and bioassay by induction of cell conformational changes of human polymorphonuclear leukocytes revealed that the parasite peptide is very similar to authentic met-enkephalin. A met-enkephalin-like peptide was also shown to be present in adult worms and in their incubation medium. Taken together, these observations demonstrate the existence of a complete opiate system in S. mansoni. We discuss its role in molecular signaling within the parasite and in host-parasite interactions.