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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Science, Ltd
    European journal of neuroscience 16 (2002), S. 0 
    ISSN: 1460-9568
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The actions of ciguatoxins from the Pacific (P-CTX-1) and Caribbean (C-CTX-1) regions were investigated in isolated parasympathetic neurons from rat intracardiac ganglia using patch-clamp recording techniques. Under current-clamp conditions, bath application of P-CTX-1 (1–10 nm) or C-CTX-1 (10–30 nm) caused a gradual depolarization that was accompanied by oscillation of the membrane potential leading to tonic action potential firing. Membrane potential oscillations were observed between −45 and −60 mV and had an amplitude of 10–20 mV and a mean frequency of 10 Hz. Oscillation frequency was temperature-dependent with a Q10 of 2.0. Membrane oscillations were temporarily inhibited by hyperpolarizing current pulses and potentiated by weak depolarizing current pulses. The amplitude of oscillations was reduced upon lowering the external Na+ concentration and inhibited by tetrodotoxin (TTX), tetracaine or Zn2+. Tetraethylammonium, 4-aminopyridine, Cs+, Cd2+, Ba2+, 1,4,4′-diothiocyanato-2,2′-stilbenedisulphonic acid (DIDS) and ouabain had no effect on the CTX-1-induced membrane depolarization and oscillations. Brevetoxin (PbTx-3, 100 nm), in contrast to CTX-1, caused a membrane depolarization that was not associated with oscillation of the membrane potential. Under voltage-clamp conditions, P-CTX-1 inhibited the peak amplitude of the voltage-dependent Na+ current and shifted the activation curve to more negative potentials, but membrane oscillations were not seen in this configuration. These results suggest that ciguatoxins cause oscillation of the membrane potential in mammalian autonomic neurons by modifying the activation and inactivation properties of a population of TTX-sensitive Na+ channels.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Science Ltd
    Molecular microbiology 53 (2004), S. 0 
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Spo0A, the response regulator protein controlling the initiation of sporulation in Bacillus, has two distinct domains, an N-terminal phosphoacceptor (or receiver) domain and a C-terminal DNA-binding (or effector) domain. The phosphoacceptor domain mediates dimerization of Spo0A on phosphorylation. A comparison of the crystal structures of phosphorylated and unphosphorylated response regulators suggests a mechanism of activation in which structural changes originating at the phosphorylatable aspartate extend to the α4β5α5 surface of the protein. In particular, the data show an important role in downstream signalling for a conserved aromatic residue (Phe-105 in Spo0A), the conformation of which alters upon phosphorylation. In this study, we have prepared a Phe-105 to Ala mutant to probe the contribution of this residue to Spo0A function. We have also made an alanine substitution of the neighbouring residue Tyr-104 that is absolutely conserved in the Spo0As of spore-forming Bacilli. The spo0A(Y104A) and spo0A(F105A) alleles severely impair sporulation in vivo. In vitro phosphorylation of the purified proteins by phosphoramidate is unaffected, but dimerization and DNA binding are abolished by the mutations. We have identified intragenic suppressor mutations of spo0A(F105A) and shown that these second-site mutations in the purified proteins restore phosphorylation-dependent dimer formation. Our data support a model in which dimerization and signal transduction between the two domains of Spo0A are mediated principally by the α4β5α5 signalling surface in the receiver domain.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Oxford [u.a.] : International Union of Crystallography (IUCr)
    Acta crystallographica 45 (1989), S. 676-678 
    ISSN: 1600-5759
    Source: Crystallography Journals Online : IUCR Backfile Archive 1948-2001
    Topics: Chemistry and Pharmacology , Geosciences , Physics
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Copenhagen : International Union of Crystallography (IUCr)
    Acta crystallographica 55 (1999), S. 1920-1922 
    ISSN: 1399-0047
    Source: Crystallography Journals Online : IUCR Backfile Archive 1948-2001
    Topics: Chemistry and Pharmacology , Geosciences , Physics
    Notes: A Bowman–Birk type trypsin/chymotrypsin inhibitor isolated from Vigna unguiculata seeds has been crystallized. Crystals were grown using the vapour-diffusion method at pH 4.0 using citrate/phosphate as a buffer and 30% saturated ammonium sulfate as precipitant. The crystals belonged to the monoclinic space group P21, with unit-cell parameters a = 32.4, b = 61.8, c = 32.9 Å, β = 114.5°. The Matthews coefficient calculated assuming two molecules in the asymmetric unit was 1.95 Å3 Da−1, which corresponds to a 37% solvent content. X-ray data were collected to 2.5 Å resolution from a flash-frozen crystal. The structure was solved using the molecular-replacement method using tracy soybean inhibitor structure (PDB entry 1pi2) as a model.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Oxford [u.a.] : International Union of Crystallography (IUCr)
    Acta crystallographica 46 (1990), S. 78-81 
    ISSN: 1600-5759
    Source: Crystallography Journals Online : IUCR Backfile Archive 1948-2001
    Topics: Chemistry and Pharmacology , Geosciences , Physics
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Naunyn-Schmiedeberg's archives of pharmacology 352 (1995), S. 236-246 
    ISSN: 1432-1912
    Keywords: Key words Ciguatoxin ; Sympathetic ganglia ; Preganglionic ; Excitability ; Sodium channel ; Synaptic excitation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  The actions of low concentrations of ciguatoxin-1 (CTX-1, 0.2–0.8 nM) in guinea-pig sympathetic ganglia have been analysed using intracellular recording techniques in vitro. The effects of CTX-1 were graded with concentration but sensitivity varied markedly between neurones in the same preparation. Other than an initial transient (∼10 min) depolarization of some ganglion cells accompanied by an increase in input resistance, passive electrical properties did not significantly differ from controls. Amplitude and threshold of action potentials evoked by depolarizing current and threshold, latency and form of the initial responses to nerve stimulation were also not affected. Exposure to CTX-1 generated marked increases in the frequency of spontaneous excitatory synaptic potentials which often occurred in bursts (15–66 Hz) of similar amplitudes. Single stimuli to incoming nerves produced repetitive synaptic responses arising from preganglionic, but not from peripheral afferent, axons. Following brief (〈5 min) exposure to CTX-1, these effects declined over 30 min but, after longer exposure (〉15 min), they persisted for several hours despite continuous washing. All activity generated by CTX-1 was significantly reduced or abolished by d-tubocurarine (10-5–10-4 M), hexamethonium (10-5 M), tetrodotoxin (10-7–10-6  M), ω-conotoxin (10-7 M), reduced Ca2+ (0.1 mM)/raised Mg2+ (10 mM), raised Ca2+ (6 mM) or raised Mg2+ (25 mM). The data suggest that CTX-1 activates preganglionic axons by modifying the voltage sensitivity of a subpopulation of Na+ channels. Effects on these unmyelinated axons occur at much lower concentrations than have been reported to affect myelinated ones. Many of the symptoms of ciguatera poisoning might be explained by activity in autonomic and perhaps other unmyelinated nerve terminals.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Naunyn-Schmiedeberg's archives of pharmacology 352 (1995), S. 236-246 
    ISSN: 1432-1912
    Keywords: Ciguatoxin ; sympathetic ganglia Preganglionic ; Excitability ; Sodium channel ; Synaptic excitation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The actions of low concentrations of ciguatoxin-1 (CTX-1, 0.2-0.8 nM) in guinea-pig sympathetic ganglia have been analysed using intracellular recording techniques in vitro. The effects of CTX-1 were graded with concentration but sensitivity varied markedly between neurones in the same preparation. Other than an initial transient (− 10 min) depolarization of some ganglion cells accompanied by an increase in input resistance, passive electrical properties did not significantly differ from controls. Amplitude and threshold of action potentials evoked by depolarizing current and threshold, latency and form of the initial responses to nerve stimulation were also not affected. Exposure to CTX-1 generated marked increases in the frequency of spontaneous excitatory synaptic potentials which often occurred in bursts (15–66 Hz) of similar amplitudes. Single stimuli to incoming nerves produced repetitive synaptic responses arising from preganglionic, but not from peripheral afferent, axons. Following brief (〈 5 min) exposure to CTX-1, these effects declined over 30 min but, after longer exposure (〉 15 min), they persisted for several hours despite continuous washing. All activity generated by CTX-1 was significantly reduced or abolished by d-tubocurarine (10−5–10−4 M), hexamethonium (10−5 M), tetrodotoxin (10−7–10−6 M), ω-conotoxin (10−7 M), reduced Ca2+ (0.1 mM)/raised Mg2+ (10 mM), raised Ca2+ (6 mM) or raised Mg2+ (25 mM). The data suggest that CTX-1 activates preganglionic axons by modifying the voltage sensitivity of a subpopulation of Na+ channels. Effects on these unmyelinated axons occur at much lower concentrations than have been reported to affect myelinated ones. Many of the symptoms of ciguatera poisoning might be explained by activity in autonomic and perhaps other unmyelinated nerve terminals.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1432-1793
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Field and laboratory experiments were used to investigate the spatial scale of benthic effects of an active nearshore produced-water (=aqueous wastes of oil and gas production) outfall on various components of recruitment in the giant kelp Macrocystis pyrifera. Results showed that discernible effects on all parameters measured were limited to areas very close to the outfall (〈50 m). Zoospore production in sporophytes transplanted to varying distances from the diffusers did not vary in a systematic way. Survival and successful reproduction (i.e., sporophyte production) of outplanted gametophytes varied significantly among experimental dates. Performance of these parameters was significantly reduced only at the site nearest the diffusers (5 m away). Poor gametophyte survival near the outfall may have resulted from exploitative competition with Beggiatoa sp. (a fast-growing filamentous marine bacteria that exploits areas high in hydrogen sulfide, an abundant constituent of the produced-water effluent) rather than from toxicity of produced water. Laboratory assays indicated that gametophyte reproduction and subsequent sporophyte production were inhibited at levels likely to occur within the near vicinity of the diffusers. Nonetheless, field data indicate that the lack of sporophyte production near the diffusers probably resulted from factors affecting gametophyte survival.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1432-1793
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Point sources of pollution (e.g. industrial and municipal outfalls) may produce ecological impacts at distant locations if pollutants affect dispersive propagules. We used laboratory experiments to determine how water-column exposure to produced water (=the aqueous fraction of oil and gas production that is typically discharged into coastal waters) influences the colonization potential of giant kelp (Macrocystis pyrifera) zoospores on the bottom. Zoospores were maintained in suspension at relatively low densities in 18-liter containers and exposed to one of five concentrations of produced water for varying amounts of time. Zoospore swimming generally decreased with increasing produced-water concentration and exposure duration; however, the specific pattern of decrease differed between experimental trials done on different dates. The effect of exposure duration on the ability of swimming zoospores to attach to plastic dishes placed on the bottom varied with produced-water concentration. Zoospores placed in produced-water concentrations of 1 and 10% showed a steady decline in their ability to attach with increased exposure; lower concentrations of produced water had no such effects. The percentage of zoospores that germinated after attachment varied tremendously with exposure duration and date of experimental trial. Zoospores that settled during the first 12 h after release had very poor rates of germination, indicative of a short precompetent period. Surprisingly, exposure of suspended zoospores to high concentrations of produced water during the first 12 h reduced this precompetent period and greatly improved germination success on the bottom. The magnitude of this enhancement, however, varied among dates. The results suggest that adverse effects of discharging produced water on planktonic zoospores of giant kelp would most likely be limited to the immediate vicinity of the outfall.
    Type of Medium: Electronic Resource
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