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1.
1.  In this commentary we discuss results obtained by a micromethod for the study of Cl– permeability across single nerve membranes from rabbit Deiters' neurons.
2.  These results showed the presence of GABAA receptors on the nerve cell membrane cytoplasmic side.
3.  We could show that these receptor complexes have a higher affinity for GABA than their extracellularly facing counterparts. Moreover, they present a phenomenon of desensitization. Another distinct property is that upon activation by GABA, they expose positive charges at their cytoplasmic mouths.
4.  We propose that these receptor complexes could functionin situ as a device for extruding Cl– anions from the nerve cell interior. This phenomenon would create an electrochemical gradient for Cl– penetration into the cell upon the action of extracellular GABA, after its presynaptic release.
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2.
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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3.
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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4.
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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5.
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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6.
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   

7.
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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8.
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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9.
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   

10.
1.  The effect of outward and inward water flows through the membrane on outward potassium currents of dialyzedHelix pomatia neurons was studied.
2.  An outward water flow increased the peak and sustained outward potassium currents and accelerated the kinetics of their activation. An inward water flow had quite opposite effects—it decreased the peak and sustained potassium currents and delayed the kinetics of their activation.
3.  The analysis of the effect of water flow on the conductance of potassium channels showed that an outward water flow increased both the potassium conductance at a given potential (gk) and the maximum potassium conductance (g k max ). An inward water flow again had the opposite effect—it decreased the potassium conductance at given potential and the maximum potassium conductance.
4.  Neither an outward nor an inward water flow significantly affected the fraction of open potassium channels at a given potential [n (V)].
5.  These data suggest that in dialyzed neurons the changes of outward potassium current during water flow through the membrane are due mainly to the changes in single-channel conductance and the time constant of current activation.
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11.
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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12.
1.  The reactions of tympanic nerve fibers ofLocusta migratoria were recorded by glass microelectrodes in the metathoracic ganglion.
2.  The units were classified by frequency-, intensity-, and directional characteristics as well as by their response pattern. The response to speciesspecific song is compared with the response to song ofEphippiger ephippiger.
3.  The physiological properties lead to a classification into three types of low-frequency neurons (characteristic frequency 3.5–4 kHz; 4kHz; 5.5–6 kHz) and one type of high-frequency neuron (12–20 kHz). This is similar to other species (Gray, 1960, Michelsen, 1971).
4.  Intensity-coding is done by sharp rising intensity characteristics and by different absolute thresholds of the units.
5.  There is a marked directional sensitivity with some differences between LF and HF units. In the low frequency range the tympanal organ seems to react as a pressure gradient receiver; for high frequencies another mechanism is discussed.
6.  No filtering of species-specific song takes place at the level of the receptor cells.
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13.
1.  Coupling mechanisms between ciliary beating and the membrane potential in Paramecium were investigated under voltage clamp applying intracellular pressure injection of cAMP, cGMP and Ca-EGTA buffer. Ciliary responses following step changes in membrane potential were recorded by high-speed video on magnetic tape.
2.  Injections of cAMP and cGMP up to millimolar concentrations caused no detectable changes in the frequency voltage relationship. A minor effect was that the ciliary reorientation towards the anterior cell end (reversal) tended to be inhibited with depolarization up to 10 mV.
3.  Injection of Ca2+ into the cell clamped at the resting potential caused a transient anteriad ciliary reorientation and a simultaneous increase in the beating frequency.
4.  Injection of EGTA (to buffer Ca2+ below 10–8 M) was ineffective in relation to frequency for several minutes. After this time, hyperpolarization- and depolarization activated frequency responses of EGTA-injected cells were increasingly inhibited. The ciliary reorientation following depolarization was not affected by EGTA.
5.  A posterior contraction of the cell diameter was noticed upon membrane hyperpolarization. The contraction coincided in time with the increase in beating frequency.
6.  The results support the view that the voltage-dependent augmentation of the ciliary beating rate is not directly mediated by an intracellular increase in either cAMP or cGMP.
7.  The role of Ca2+ as intracellular messenger in the ciliary and somatic compartments is discussed.
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14.
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   

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.
Metabolic characteristics of experimental hepatoma cells include elevated rates of glycolysis and lipid synthesis. However, pyruvate derived from glucose is not redily oxidized, and the source of acetly CoA for lipid synthesis in As-39D cells has not been characterized. In this study ketone bodies were examined as a possible source of acetyl CoA in AS-30D hepatoma cells.The major findings were:
1.  Acetoacetate was utilized by AS-30D cells, with14C-lipid and14CO2 as major products of [3-14C] acetoacetate.
2.  Lipid synthesis from acetoacetate was dependent on the presence of glucose in the medium.
3.  Acetoacetate supported rapid respiration by AS-30D mitochondria in the presence of 0.1 mM malate.
4.  Succinly CoA acetoacetyl CoA transferase activity in AS-30D mitochondria was approximately 40 fold greater than that found in rat liver mitochondria.
5.  Addition of acetoacetate, but not -hydroxybutyrate decreased conversion of [1-14C] acetate to14CO2, presumably by diluting the specific radioactivity of the acetyl CoA derived from the acetate tracer.
6.  In the presence of glucose, approximately one fourth of acetoacetate utilized was converted to lipid. This result is consistent with elevated lipogenesis postulated by the truncated TCA cycle hypothesis. These data demonstrate for the first time the flux of acetoacetate carbon to lipid and CO2 in hepatoma cells and suggest that increases in the ambient concentration of acetoacetate, occurring in fasting or malignant cachexia, could produce increases in the utilization of this ketone body by hepatoma cells containing 3-oxyacid CoA transferase activity.
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17.
1.  Transepithelial potential differences (PDte) and acidification rates of the bath chamber were measured on isolated perfused posterior gills of the fiddler crabUca tangeri adapted to dilute seawater.
2.  The PDte decreased to almost zero when Na+ was substituted by choline or when ouabain was added to the perfusion saline in high concentrations (10 mmol·l–1). Thus, the rheogenic NaCl-transport across the gill epithelium seems to be totally Na+-dependent.
3.  When Cl– was replaced by gluconate, a bath positive PDte occurred which was insensitive to ouabain. This PDte could also be observed when, in addition to Cl– removal, Na+ was replaced by TMA+.
4.  Bath acidification under normal conditions could be abolished by ouabain, indicating that there is H+ excretion via electrically silent Na+/H+ exchange. In contrast, bath acidification under Cl–-free conditions is only partially blocked by ouabain.
5.  It is concluded that under Cl–-free conditions a rheogenic H+-pump in the apical membrane is responsible for the ouabain-insensitive bath acidification as well as for the PDte.
Parts of this study had been published earlier in abstract form  相似文献   

18.
Oncostatin M, a multifunctional cytokine   总被引：6，自引：0，他引：6  
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19.
1.  Muscles of the posterior cardiac plate (pcp) and pyloric regions in the stomach of Squilla are innervated by motoneurons located in the stomatogastric ganglion (STG). The pattern of innervation of various muscles in these regions was determined using electrophysiological methods.
2.  The dilator muscles are singly or doubly innervated by the pyloric dilator neurons (PDs). The constrictor muscles are singly or doubly innervated by the pcp neuron (PCP) or the pyloric neurons (PYs). These muscles are sequentially activated by pcp-pyloric motor outputs produced by the PCP, PY, and PD. All muscles can generate an all-or-nothing spike.
3.  The constrictor muscles generate spikes followed by depolarizing afterpotentials which lead to a sustained depolarization with repetitive spikes. The PYs can entrain rhythmic spike discharges of these muscles.
4.  The spike of muscles remains unchanged by bath application of tetrodotoxin (10-7 M) to suppress neuronal impulse activities, but it is blocked by Mn2+ (10 mM).
5.  The constrictor muscle isolated from the STG displays an endogenous property of spontaneous membrane oscillation that produces a train of spikes. Brief depolarizing or hyperpolarizing stimuli can trigger or terminate an oscillatory potential, respectively, and reset the subsequent rhythm.
6.  The possible functions of myogenicity under the control of discharges of motoneurons in the pyloric constrictor neuromuscular system are discussed.
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20.
1.  The self-adapting effects of chemical backgrounds on the response of primary chemoreceptor cells to superimposed stimuli were studied using lobster (Homarus americanus) NH4 receptor cells.
2.  These receptors responded for several seconds to the onset of the backgrounds, and then returned to their initial level of spontaneous activity (usually zero). The strongest response always occurred only during the steepest concentration change; the response then decayed back to zero or to the earlier spontaneous firing level, while the background concentration was still rising, and remained silent during the entire time that the background was maintained constant (20–30 min) (Fig. 2).
3.  Exposure to constant self-adapting backgrounds eliminated the responses of NH4 receptor cells to stimuli of concentration lower than the background, and reduced the responses to all higher stimulus concentrations tested by a nearly equal amount. This resulted in a parallel shift of the stimulus-response function to the right along the abscissa (Figs. 3 and 4).
4.  Since the response threshold was completely re-set by adaptation to backgrounds, NH4 receptors seem to function mostly as detectors of relative rather than absolute stimulus intensity across their entire dynamic range: the response to a given stimulus-to-background ratio remained the same over 3 log step increases of background concentration (Fig. 6).
5.  As in other sensory modalities, a parallel shift of response functions appears to be an important property of chemoreceptor cells, allowing for this sensory system to function over a wider stimulus intensity range than the instantaneous dynamic range of individual receptor cells.
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