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1.
1.  Properties of the membrane currents ofDidinium nasutum have been investigated under voltage clamp in different solutions and after deciliation.
2.  Theearly transient Ca2+ inward current activates in a voltage-dependent manner. Inactivation is both Ca2+ -dependent and voltage-dependent.
3.  Alate Ca2+ current rises with time to peak > 50 ms and decays in the order of seconds.
4.  Activation and inactivation of the late Ca2+ current is voltage-dependent.
5.  The delayed outward current is activated by voltage. The kinetics of this K+ current, but not its amplitude, are enhanced in the presence of intracellular EGTA.
6.  The two voltage-dependent Ca2+ channels are located in the cilia, whereas all K+ channels are restricted to the somatic membrane.
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2.
1.  The electrogenic Na-K pump activity was studied in thePlotosus electroreceptor. Single ampullae (sensory epithelium) were dissected free, and mounted over an air gap at the ampullary duct, to electrically isolate the receptor activity for current- and voltage-clamp experiments.
2.  After equilibration in K-free saline, the bathing medium around the ampulla was exchanged to test saline. Under current-clamp, 4 mM K saline induced reversible hyperpolarization of about 13 mV. Under voltage-clamp, the 4 mM saline induced an outward current of about 10 nA.
3.  Both responses were suppressed by ouabain, with a half suppression at 1.5×10–7 M for current, which suggested involvement of ionic pumps. In the K-free saline, ouabain alone induced no response.
4.  The current response to high K saline was always outward, and was dependent on K concentration. In the presence of ouabain, high K saline as high as 40 mM, ten times the standard K, induced no inward current, which suggested little contribution of K conductance to the responses.
5.  The outward current responses were induced, also dose-dependently, by various alkali metal cations. The apparent dissociation constants were 6.8, 1.2, 21.7, and 85.1 mM for K, Rb, Cs, and Li, suggesting their potency in the order of Rb>K> Cs>Li.
6.  Thus, the hyperpolarization and outward current responses were due to an electrogenic NaK pump in the basal face of the sensory epithelium. The Na-K pump seems to be responsible, to a certain extent, for maintenance of the negative DC potential in the standard 4 mM K saline, which supplies an outward current to bias the sensory epithelium in situ.
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3.
1.  At ambient temperatures (T a) between 39 and 43°C, specimens of the waterproof treefrogChiromantis xerampelina, resting quietly in wind tunnels, adjust rates of evaporative water loss (EWL), maintaining body temperature (T b) 2–4°C belowT a.
2.  Brain heating and cooling, respectively, increased and decreased steady-state rates of thermoregulatory evaporative water loss (EWL), driving negative feedback changes inT b.
3.  Continuous infusions of epinephrine resulted in specific, dose-dependent, saturable increases in EWL; isoproterenol was more potent than epinephrine, which was more potent than phenylephrine. Tyramine injection also stimulated EWL.
4.  Non-specific increases in EWL stimulated by injections of cholinergic agonists were weakly antagonized by atropine, but thermally induced EWL, as well as adrenergically and cholinergically stimulated increases in EWL, were abolished by beta-adrenergic antagonists.
5.  Sweating decreased andT b increased during ganglionic blockade.
6.  The observations suggest thatC. xerampelina controls thermoregulatory EWL by modulating the sympathetic nervous outflow stimulating beta-adrenergic receptors on cutaneous mucous glands.
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4.
1.  The diffusive hydrogen conductance of chicken eggshell compound membrane was measured in situ on day 16 of incubation, in a direction parallel to the shell and the chorioallantois (lateral conductance). A value of 3.9 mmol d–1 kPa–1 was obtained through a ring 13.29 cm in circumference, 0.0076 cm thick and 0.3 cm long.
2.  Lateral hydrogen conductance for 1 mm2 of shell membrane 76 m thick is 30 times the conductance of one pore serving the same area.
3.  Lateral conductance for H2 is not significantly influenced by chorioallantoic perfusion.
4.  Oxygen consumption change due to partial covering of the hen eggshell indicates that there is a significant resistance to lateral diffusion of oxygen under the shell toward the covered area.
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5.
1.  The effects of three barbiturates and the local anesthetic procaine on the ion channel function of mouse nicotinic acetylcholine receptor (nAChR) muscle subtype expressed inXenopus laevis oocytes were examined by whole-cell voltage-clamp technique.
2.  A concentration-response curve for the specific nicotinic agonist dimethylphenylpiperazinium iodide (DMPP) was first determined. This agonist produced increasing whole-cell currents up to a concentration of 100µM (EC50 = 13µM), then decreased responses at higher concentrations.
3.  The barbiturates (amobarbital, secobarbital, pentobarbital) and procaine produced reversible inhibition of DMPP-induced currents at clinically used concentrations. The two classes of drugs differed in the voltage dependence of the inhibition: procaine-induced inhibition was increased at more negative transmembrane holding potentials (-90 vs. -45 mV); whereas amobarbital-induced inhibition did not vary at different transmembrane potentials.
4.  Mutant forms of the nAChR, containing single amino acid changes in the M2 regions of and subunits, showed increased sensitivity to procaine but no change in sensitivity to amobarbital-induced inhibition.
5.  These electrophysiologic studies provide further evidence that barbiturates and local anesthetics produce inhibition of the nAChR at different sites.
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6.
1.  The actions of GABA on three classes of visual interneurons in crayfish, Procambarus clarkii, medulla externa are examined. The effect of GABA on the visual response is compared to GABA's action on agonist-elicited responses purported to mediate the visual response.
2.  GABA produces a shunting type of inhibition in medullary amacrine cells which is associated with a small depolarization (Figs. 2, 3), a large increase in input conductance (Gn) and a reversal potential close to rest (Fig. 4). GABA is a potent antagonist to the depolarizing action of acetylcholine (ACh) (Fig. 5).
3.  GABA depolarizes dimming fibers (Fig. 2), and the response is mediated by an increase in Gn (Fig. 6). GABA antagonizes the light-elicited IPSP and the hyperpolarizing action of ACh (Fig. 7).
4.  Sustaining fibers (SF) do not appear to have GABA receptors but GABA inhibits the excitatory visual input pathway to the SFs (Fig. 8). Conversely, the GABA antagonist, bicuculline, potentiates the SF light response (Fig. 9).
5.  GABA has at least three different modes of antagonist action in the medulla: i) Increased conductance and depolarization in dimming fibers and medullary amacrine neurons; ii) Decreased chloride conductance in tangential cells; and iii) An inhibitory action on the visual pathway which drives SFs.
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7.
1.  Up to 9 kHz, the tympanal membrane of the grasshopper Chorthippus biguttulus responds with equal sensitivity at the attachment sites of the low and the high-frequency receptors; at the latter site it is also particularly sensitive between 10 and 20 kHz.
2.  The frequency spectra of the songs of both sexes exhibit maxima at 7–8 kHz, to which the membrane is well matched. In the high-frequency region, where the male songs have a peak at 30 kHz, there is no corresponding maximum in the membrane oscillation.
3.  Because the tympanal membrane is immediately adjacent to air sacs in the tracheal system, it is deflected inward and outward by as much as 80 m during the respiratory cycle.
4.  Measurements by laser vibrometry show that acoustically induced membrane oscillations are attenuated severely due to the respiratory displacement of the membrane for frequencies up to 10–12 kHz. By contrast, at higher frequencies the membrane sensitivity is doubled or tripled.
5.  As a result of these membrane effects, the discharge in the tympanal nerve was profoundly reduced in the low-frequency range, whereas above 11 kHz there was a marked increase. This modulation of auditory sensitivity affects the animals' ability to detect conspecific songs.
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8.
1.  The effects of potassium channel blockade on afferent axons and terminal regions in frog dorsal roots and spinal cords, respectively, were investigatedin vitro.
2.  A condition-test (C-T) protocol was used to assess the population relative refractory period. Characteristics of main axons were evaluated by stimulation at the proximal end of transected dorsal roots (DR). Characteristics of terminal regions were tested by stimulation at the base of the dorsal horn (DH).
3.  DH recovery of excitability was delayed by low concentrations of 4-aminopyridine (4-AP) and tetraethylammonium (TEA) alone or combined. The same treatments did not affect recovery to DR stimulation.
4.  DH recovery of excitability was not delayed by solutions suppressing terminal calcium influx.
5.  We conclude that sensitivity of the relative refractory period to potassium channel blocking agents differs between main axons and axon terminal regions. This may indicate differences between axon terminals and main axons in the mechanism of action potential repolarization.
6.  We hypothesize that rapid action potential repolarization by pharmacologically sensitive potassium channels in presynaptic terminal regions keeps terminal action potentials short. Terminal action potential brevity would limit calcium influx, thus preventing terminal calcium overload but contributing to transmission failures at spinal synapses.
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9.
1.  Three cyclic diterpenoids isolated from gorgonians of theEunicea genus and characterized as eupalmerin acetate (EUAC), 12,13-bisepieupalmerin (BEEP), and eunicin (EUNI) were found to be pharmacologically active on the nicotinic acetylcholine receptor (AChR).
2.  The receptor from the BC3H-1 muscle cell line was expressed inXenopus laevis oocytes and studied with a two-electrode voltage clamp apparatus.
3.  All three compounds reversibly inhibited ACh-induced currents, with IC50's from 6 to 35µM. ACh dose-response curves suggested that his inhibition was noncompetitive. The cembranoids also increased the rate of receptor desensitization.
4.  Radioligand-binding studies using AChR-rich membranes fromTorpedo electric organ indicated that all three cembranoids inhibited high-affinity [3H]phencyclidine binding, with IC50's of 0.8, 11.6, and 63.8µM for EUNI, EUAC, and BEEP, respectively. The cembranoids at a 100µM concentration did not inhibit [-125I]bungarotoxin binding to either membrane-bound or solubilized AChR.
5.  It is concluded that these compounds act as noncompetitive inhibitors of peripheral AChR.
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10.
1.  Seasonal changes in the half-times (0.5) for calcium influx and outflux in bone, skin and scales, and soft-tissue compartments ofF. kansae were measured.
2.  The size of diffusible calcium pools in the three compartments was estimated from the ratio of the loading and unloading half-times. Both the bone and skin and scales compartments contain diffusible calcium pools which increase in size during the summer months.
3.  The calcium in the soft-tissue compartment is all diffusible, and the kinetics indicate that this compartment receives calcium from the other two compartments.
4.  Hypophysectomy reduced the rate of calcium turnover in summer animals to the levels measured in intact winter animals. Such treatment also affected the distribution of radiocalcium within the animal.
5.  It is concluded that acellular bone is a metabolically active tissue.
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11.
1.  Normal activity in bilateral pairs of heart interneurons, from ganglia 3 or 4, in the medicinal leech (Hirudo medicinalis) is antiphasic due to their reciprocally inhibitory connections. However, Ca+-free Co+-containing salines lead to synchronous oscillations in these neurons.
2.  Internal TEA+ allows expression of full plateaus during Co++ induced oscillations in heart interneurons; these plateaus are not blocked by Cs+. Similar plateaus are also observed with internal TEA+ alone, but under these conditions activity in heart interneurons from ganglia 3 or 4 is antiphasic.
3.  Plateaus in heart interneurons induced by Co++ and internal TEA+ involve a conductance increase.
4.  A voltage-dependent inward current, IP, showing little inactivation, was isolated using single-electrode voltageclamp in heart interneurons. This current is carried at least in part by Na+; the current is reduced when external Na+ is reduced and is carried by Li+ when substituted for Na+.
5.  Calcium channel blockers such as La3+ and Co++ block neither the TEA+ induced plateaus nor IP, suggesting that Na+ is not using Ca++ channels. Moreover, IP is enhanced by Ca++-free Co++-containing salines. Thus, IP is correlated with the TEA+- and Co++-induced plateau behavior.
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12.
Conclusions  
(1)  The aminoesters inhibit glucose-stimulated proton extrusion by yeast cells.
(2)  The inhibitory activity depends on aliphatic carbon chain length.
(3)  The inhibition of proton extrusion is concentration-dependent.
(4)  The aminoesters stimulate quinacrine accumulation in vacuoles of yeast cells so they should possess affinities for lysosomes.
This work was supported byKBN grant no. 7 A203 013 07.  相似文献   

13.
1.  We have obtained a cDNA clone encoding a human retinal D2 dopamine receptor.
2.  The longest open reading frame (1242 bp) of this clone encodes a protein of 414 amino acids having a predicted molecular weight of 47,000 and a transmembrane topology similar to that of other G protein-coupled receptors.
3.  Transient transfection of COS-7 cells with an expression vector containing the clone resulted in expression of a protein possessing a pharmacological profile similar to that of the D2 dopamine receptor found in striatum and retina.
4.  Northern blot analysis indicated that, in rat brain and retina, the mRNA for this receptor was 2.9 kb in size.
5.  In situ hybridization was performed to examine the distribution of the mRNA for this receptor in human retina. Specific hybridization was detected in both the inner and the outer nuclear layers.
6.  These findings are consistent with prior physiological and autoradiographic studies describing the localization of D2 dopamine receptors in vertebrate retinas. Our observations suggest that photoreceptors as well as cells in the inner nuclear layer of human retinas may express the mRNA for this D2 dopamine receptor.
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14.
3DFS is a 3D flexible searching system for lead discovery. Version 1.0 of 3DFS was published recently (Wang, T.; Zhou, J. J. Chem. Inf. Comput. Sci., 1998, 38, 71–77). Here version 1.2 represents a substantial improvement over version 1.0. There are six major changes in version 1.2 compared to version 1.0.
1.  A new rule of aromatic ring recognition.
2.  The inclusion of multiple-type atoms and chains in queries.
3.  The inclusion of more spatial constraints, especially the directions of lone pairs.
4.  The improvement of the query file format.
5.  The addition of genetic search for flexible search.
6.  An output option for generating MOLfiles of hits.
Besides the above, this paper supplies:
1.  More query examples.
2.  A comparison between genetic search and Powell optimization.
3.  More detailed comparison between 3DFS and Chem-X.
4.  A preliminary application of 3DFS to K+ channel opener studies.
Supplementary material to this paper is available in electronic form at http://dx.doi.org/10.1007/s0089490050231  相似文献   

15.
Müller  D. G.  Frenzer  K. 《Hydrobiologia》1993,(1):37-44
Culture studies with healthy and virus-infected isolates of Ectocarpus siliculosus, Feldmannia simplex and F. irregularis gave the following results:
–  Virus particles are produced in deformed reproductive organs (sporangia or gametangia) of the hosts and are released into the surrounding seawater.
–  Their infective potential is lost after several days of storage under laboratory conditions.
–  New infections occur when gametes or spores of the host get in contact with virus particles. The virus genome enters all cells of the developing new plant via mitosis.
–  Virus expression is variable, and in many cases the viability of the host is not impaired. Infected host plants may be partly fertile and pass the infection to their daughter plants.
–  Meiosis of the host can eliminate the virus genome and generate healthy progeny.
–  The genome of the Ectocarpus virus consists of dsDNA. Meiotic segregation patterns suggest an intimate association between virus genome and host chromosomes.
–  An extra-generic host range has been demonstrated for the Ectocarpus virus.
–  Field observations suggest that virus infections in ectocarpalean algae occur on all coasts of the world, and many or all Ectocarpus and Feldmannia populations are subject to contact with virus genomes.
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16.
1.  Rates of oxygen consumption were measured during locomotion in five species of marsupials of the family Dasyuridae. The body weights of the animals ranged between 0.15 and 1.12 kilograms.
2.  The rate of change of power input with speed was generally lower than equivalent eutherian values. The extrapolation to zero speed was consistently a higher multiple of resting metabolic levels than found in eutherians.
3.  The minimum cost of locomotion (M run) as a function of body mass (wt) is described by the equationM run=4.75 wt–0.34. The exponent is similar to that described for eutherians and reptiles, but the constant term is significantly lower.
4.  Metabolic scope in these animals is similar over the size range used and may be greater than in eutherians.
5.  Heat dissipation during locomotion has been partitioned into evaporative and non-evaporative routes. Storage of heat during locomotion was never more than fifty per cent of total production.
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17.
Twelve of the main European LCA software packages currently available are examined wirh the aim of establishing which are the most appropriate for LCAs on industrial processes. The packages performances are assessed in terms of
–  • Volume of Data
–  • WindowsTM environment
–  • Network Capabilities
–  • Impact Assessment
–  • Graphical representation of the inventory results
–  • Sensitivity analysis
–  • Units
–  • Cost
–  • User Support
–  • Flow Diagrams
–  • Burdens allocation
–  • Transparency of data
–  • Input & output parameters
–  • Demo version
–  • Quality of data
The review concludes with a Specification Table which summarises the facilities available on each software package. The general conclusion from this study is that for industrially based LCAs, there are four packages which may offer advantages over the rest. These are The Boustead Model, The Ecobilan Group’s TEAM™, PEMS 3.0 and SimaPro 3.1.  相似文献   

18.
1.  Authentic human acetylcholinesterase (AChE) was expressed inEscherichia coli under regulation of the constitutivedeo promoter or the thermoinduciblePL promoter.
2.  To facilitate expression in the prokaryotic system, recombinant human AChE (rhAChE) cDNA was modified at the N terminus by oligonucleotide substitutions in order to replace some of the GC-rich regions by AT. These modifications did not alter the amino acid sequence but resulted in ample production of the protein.
3.  rhAChE accumulated in the cells and reached a level of 10% of total bacterial proteins. A partially purified inactive recombinant protein was recovered from inclusion bodies.
4.  Active rhAChE was obtained after solubilization, folding, and oxidation, although the recovery of the active enzyme was low. A 20- to 40-fold increase in enzymatically active rhAChE was achieved by replacing Cys580 by serine.
5.  The recombinant enzyme analogue was indistinguishable from native AChE isolated from erythrocytes in terms of substrate specificity and inhibitor selectivity.
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19.
1.  Paramecium bursaria was stimulated by a light spot of 10–15 m diameter, and the photosensitive site was searched by recording responses in swimming behavior and in membrane potential.
2.  Local stimulation to the anterior half of the cell caused an avoiding response.
3.  Stimulation to the cells deciliated by ethanol treatment elicited a depolarization of the membrane potential.
4.  Local stimulation to the anteroventral portion elicited a depolarization, but stimulation to the dorsal side induced no change in the membrane potential.
5.  The action spectrum of depolarization elicited by local stimulation to the anteroventral surface showed two main peaks at 420 nm and 560 nm, corresponding to those of light stimulation of the whole cell.
6.  It is concluded that a photosensitive site exists on the anteroventral surface ofParamecium, in particular within the oral groove of the cell. This local photosensitivity is discussed with respect to the mating reaction.
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20.
1.  Direct contact between intra-epithelial nerve endings and ciliated cells was observed in frog (Rana pipiens) palate epithelium.
2.  Electrical stimulation of the palatine nerve to the explant or the explant culture induced an increase in ciliary beat frequency in explant and outgrowth cells.
3.  Atropine inhibited electrically stimulated ciliary beat frequency increase in the explant and outgrowth cells.
4.  Gap junctional intercellular communication appears to be involved in the propagation of stimulated ciliary beat frequency increase from innervated to non-innervated ciliated cells.
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