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Xenopus neural crest cell migration in an applied electrical field.

Xenopus neural crest cells migrated toward the cathode in an applied electrical field of 10 mV/mm or greater. This behavior was observed in relatively isolated cells, as well as in groups of neural crest cells; however, the velocity of directed migration usually declined when a cell made close contact with other cells. Melanocytes with a full complement of evenly distributed melanosomes did not migrate of their own accord, but could be distorted and pulled by unpigmented neural crest cells. Incompletely differentiated melanocytes and melanocytes with aggregated melanosomes displayed the same behavior as undifferentiated neural crest cells, that is, migration toward the cathode. An electrical field of 10 mV/mm corresponded to a voltage drop of less than 1 mV across the diameter of each cell; the outer epithelium of Xenopus embryos drives an endogenous transembryonic current that may produce voltage gradients of nearly this magnitude within high-resistance regions of the embryo. We, therefore, propose that electrical current produced by the skin battery present in these embryos may act as a vector to guide neural crest migration.

Animals↗

Molecular heterogeneity of lactic dehydrogenase in avian malaria (Plasmodium lophurae).

Lactic dehydrogenase activity increased in direct proportion to the degree of parasitization in synchronous infections of duck erythrocytes. Deviations from this linearity could be accounted for on the basis of the developmental stage of the parasite. Erythrocyte-free P. lophurae showed activities which averaged 3 times that of uninfected erythrocytes, whereas infected erythrocytes had intermediate values. In addition, a patent infection was generally reflected by an increase in the lactic dehydrogenase activity in the plasma, but no direct correlation with parasitemia was established. Molecular heterogeneity of the enzyme was determined on the basis of kinetic data and electrophoretic isolation on a starch block. The uninfected red blood cell showed a major anodal and a minor cathodal peak of lactic dehydrogenase activity, and was further characterized by a kinetic constant representing a high pH optimum with low concentrations of substrate. Isolated P. lophurae had a single, cathodal peak of activity dissimilar from that of the uninfected erythrocyte, and a kinetic constant describing a low pH optimum with a high concentration of substrate. Infected erythrocytes showed a combination of these electrophoretic entities and an intermediate range of kinetic constants. The data indicate that the avian malaria parasite P. lophurae contains a lactic dehydrogenase qualitatively dissimilar from that of its host cell, and the increased enzymatic activity of infected erythrocytes is a result of the enzyme content of the growing parasite added to that of the red blood cell. It is suggested that the LDH of the parasite has a physiological advantage under those conditions which prevail inside the red blood cell.

Animals↗

Macrophage plasma membrane. II. Studies on synthesis and turnover of protein constituents.

Rabbit alveolar macrophages were incubated in vitro with radioactive protein precursors. Plasma membranes were isolated from these cells, dissolved in phenol-urea-acetic acid, and separated by acrylamide gel electrophoresis. (3)H-leucine was rapidly incorporated into membrane protein. The rate of labeling with (3)H-leucine was markedly different from one protein band to another, indicating heterogeneous or multistep synthesis and assembly of proteins in the alveolar macrophage plasma membrane. Cells incubated with (3)H-choline incorporated this compound into membrane lecithin. On gel electrophoresis the label derived from choline was located in the two bands migrating most rapidly towards the cathode. Studies on cells incubated with (3)H-glucosamine revealed incorporation of label into two protein bands, one located near the origin and the other migrating rapidly towards the cathode. The in vitro techniques were also employed for pulse-chase studies to gain information on rate of turnover of macrophage plasma membrane proteins. This turnover rate was rapid, with a half-life of approximately 8 hr. The radioactivity disappeared from the several protein bands at the same rate, suggesting bulk removal of membrane rather than catabolism of the individual proteins in situ. Endocytosis seems a likely mechanism to account for a major part of the plasma membrane removal. Studies on the protein components of phagolysosomal membranes from cells which had been labeled with (3)H-leucine revealed the presence of all of the major labeled protein bands characteristic of the plasma membrane except one, thus confirming the bulk interiorization of large segments or units of plasma membrane by endocytic processes.

Animals↗

Prolonged action potentials from single nodes of Ranvier.

The duration of action potentials from single nodes of Ranvier can be increased by several methods. Extraction of water from the node (e.g. by 2 to 3 M glycerin) causes increased durations up to 1000 msec. 1 to 5 min. after application of the glycerin the duration of the action potential again decreases to the normal value. Another type of prolonged action potential can be observed in solutions which contain K or Rb ions at concentrations between 50 mM and 2 M. The nodes respond only if the resting potential is restored by anodal current. The kinetics of these action potentials is slightly different. Their maximal durations are longer (up to 10 sec.). Like the normal action potential, they are initiated by cathodal make or anodal break. They also occur in external solutions which contain no sodium. The same type of action potentials as in KCl is found when the node is depolarized for some time (15 to 90 sec., 100 to 200 mv.) and is then stimulated by cathodal current. These action potentials require no K or Na ions in the external medium. Their maximal duration increases with the strength and duration of the preceding depolarization. The possible origin of the action potentials in KCl and after depolarization, and their relation to the normal action potentials and the negative after-potential are discussed.

Action Potentials↗

EVIDENCE FOR ANION-PERMSELECTIVE MEMBRANE IN CRAYFISH MUSCLE FIBERS AND ITS POSSIBLE ROLE IN EXCITATION-CONTRACTION COUPLING.

Under certain conditions only, isolated crayfish skeletal muscle fibers change in appearance, becoming grainy, darkening, and seemingly losing their striations. These changes result from development of large vesicles on both sides of the Z-line. The longitudinal sarcoplasmic reticulum remains unaffected. The vesicles are due to swelling of a transverse tubular system (TTS) which is presumably homologous with the T-system tubules of other muscle fibers. The vesiculations occur during efflux of water or on reducing external K or Cl, but only when KCl can leave the fiber. They never result from osmotic, ionic, or electrical changes when KCl cannot leave. Inward currents, applied through a KCl-filled intracellular cathode, also cause the vesiculations. These are not produced when the cathode is filled with K-propionate, nor by outward or longitudinal currents. Thus the transverse tubules swell only when Cl leaves the cell. Accordingly, their membrane is largely or exclusively anion-permselective. These findings also indicate that the TTS forms part of a current loop, connecting with the exterior of the fiber probably through radial tubules (RT) possessing membrane of low conductivity. Thus, part of the current flowing inward across the sarcolemma during activity can return to the exterior through the membrane of the TTS. The structure and properties of the latter offer the possibility for an efficient electrical mechanism to initiate excitation-contraction coupling.

Animals↗

THE EFFECT OF ELECTROTONUS ON THE OLFACTORY EPITHELIUM.

The effect of electrotonus on the slow potential of the olfactory epithelium of the frog was studied. The "on"-slow potential induced by a general odor like amyl acetate increased its magnitude in accordance with increase of anodal current, while it decreased its magnitude with increase of cathodal current. Similar relations were also found in the case of the vapors of organic solvents like ethyl ether of low concentrations. Conversely, the on-slow potential induced by the vapors of organic solvents of high concentration decreased its magnitude in accordance with the increase of anodal current, while it increased its magnitude with the increase of cathodal current. The "off"-slow potential induced by the vapors of organic solvents of high concentration showed a potential change under the action of electrotonic currents which is similar to the change of the on-slow potential induced by general odors. It was concluded that there are two receptive processes in the olfactory cell. One is an ordinary excitatory process which produces an electronegative slow potential in response to general odors. The other is a process of a different kind which is activated only by the vapor of an organic solvent of high concentration and which shows an entirely opposite reaction from that generally found in excitable tissues when an electrotonic current is applied.

Acetates↗

Taste transduction mechanism: similar effects of various modifications of gustatory receptors on neural responses to chemical and electrical stimulation in the frog.

Responses in the frog glossopharyngeal nerve induced by electrical stimulation of the tongue were compared with those induced by chemical stimuli under various conditions. (a) Anodal stimulation induced much larger responses than cathodal stimulation, and anodal stimulation of the tongue adapted to 5 mM MgCl2 produced much larger responses than stimulation with the tongue adapted to 10 mM NaCl at equal current intensities, as chemical stimulation with MgCl2 produced much larger responses than stimulation with NaCl at equal concentration. (b) The enhansive and suppressive effects of 8-anilino-1-naphthalenesulfonate, NiCl2, and uranyl acetate on the responses to anodal current were similar to those on the responses to chemical stimulation. (c) Anodal stimulation of the tongue adapted to 50 mM CaCl2 resulted in a large response, whereas application of 1 M CaCl2 to the tongue adapted to 50 mM CaCl2 produced only a small response. This, together with theoretical considerations, suggested that the accumulation of salts on the tongue surface is not the cause of the generation of the response to anodal current. (d) Cathodal current suppressed the responses induced by 1 mM CaCl2, 0.3 M ethanol, and distilled water. (e) The addition of EGTA or Ca-channel blockers (CdCl2 and verapamil) to the perfusing solution of the lingual artery reversibly suppressed both the responses to chemical stimulus (NaCl) and to anodal current with 10 mM NaCl. (f) We assume from the results obtained that electrical current from the microvillus membrane of a taste cell to the synaptic area supplied by anodal stimulation or induced by chemical stimulation activates the voltage-dependent Ca channel at the synaptic area.

Anilino Naphthalenesulfonates↗

Proton-induced characteristic x-rays: a versatile source of ultrasoft x-rays for biological and biochemical investigations.

Very low energy ( ultrasoft ) x-rays of 0.3-5 keV have provided a unique tool for investigation of mechanisms of radiation action, especially with respect to the energy and spatial properties of critical radiation damage in mammalian cells. Experimental investigations to date have been partially limited by the availability and characteristics of suitable ultrasoft x-ray sources. The suitability of small electrostatic proton accelerators, such as exist in many laboratories, have been investigated as a means of producing a secondary beam of ultrasoft x-rays suitable for irradiation of biological and biochemical systems. Results are presented on the physical characteristic of carbon K (0.28 keV) and aluminium K (1.5 keV) ultrasoft x-ray beams produced by bombardment of solid targets of carbon and aluminium with protons of energies up to 750 kV and currents up to 500 microA. These characteristics are compared with those of a cold cathode discharge ultrasoft x-ray tube previously used for mammalian cell investigations. It is seen that the proton accelerator produces much more versatile beams of characteristic ultrasoft x-rays which greatly extend the scope for future experiments on mammalian cells, micro-organisms and biochemical systems. Nevertheless there are situations in which the cold-cathode discharge tube will remain the source of choice and there are other situations, requiring for example energies between characteristic lines, where the greatly more complex synchrotron radiation sources are required.

Animals↗

The use of CVD diamond to produce carbon K ultrasoft x-rays.

Carbon K ultrasoft x-rays (278 eV) interact with biological material producing random, isolated tracks of single electrons with a range < 7 nm (cf width of DNA helix approximately2 nm). The electron tracks produced by these ultrasoft x-rays are similar to the numerous secondary electrons and 'track ends' produced by essentially all ionizing radiations. They therefore provide a unique tool in the study of mechanisms of radiation action in the cell. Conventional carbon targets used with the Medical Research Councils cold-cathode discharge tubes have a limited lifetime of 40-50 min. We have investigated the use of thin, freestanding diamond films produced using chemical vapour deposition techniques as a new transmission target for a cold-cathode tube. We present here a novel use of diamond to produce CK ultrasoft x-rays for irradiation purposes. The system described produces an entrance absorbed dose rate to attached cells of approximately 0.2 Gy min(-1) which due to the long target lifetime is usable for low dose-rate CK x-ray studies.

Diamond↗

Prototype heel effect compensation filter for cone-beam CT.

The prototype cone-beam CT (CBCT) has a larger beam width than the conventional multi-detector row CT (MDCT). This causes a non-uniform angular distribution of the x-ray beam intensity known as the heel effect. Scan conditions for CBCT tube current are adjusted on the anode side to obtain an acceptable clinical image quality. However, as the dose is greater on the cathode side than on the anode side, the signal-to-noise ratio on the cathode side is excessively high, resulting in an unnecessary dose amount. To compensate for the heel effect, we developed a heel effect compensation (HEC) filter. The HEC filter rendered the dose distribution uniform and reduced the dose by an average of 25% for free air and by 20% for CTDI phantoms compared to doses with the conventional filter. In addition, its effect in rendering the effective energy uniform resulted in an improvement in image quality. This new HEC filter may be useful in cone-beam CT studies.

Equipment Design↗

Computerized hematology: operation of a high-volume hematology laboratory.

Operation of automated hematology testing, on-line to a laboratory-dedicated computer, is described. The computer stores, retrieves and monitors the results of two Model S Sr. Coulter Counters, three Technicon platelet counters, one Electra 600-D, and six leukocyte differential consoles, which are interphased to the computer. All other hematology tests are batch-entered via the keyboard of cathode-ray tubes. The computer generates specimen labels, worksheets, and lists of incomplete tests, monitors all on-line instruments, and performs all the calculations used in the procedures of quality control. Results are available instantaneously on cathod-ray tubes strategically located in patient-related areas throughout the institution. These can be obtained as all of the results for a given day or as cumulative summaries or histogram-type plots of results for a given day.

Autoanalysis↗

The polarographic measurement of halothane.

The polarographic reduction of halothane on silver cathodes was studied. Silver catalyses the reduction of halothane: two electrons are exchanged in the reduction process. Experiments using a rotating disc electrode showed a linear relationship between observed current and halothane concentration. A Silastic membrane-covered polarographic electrode was constructed using a silver cathode which gave a good linear response to changes in halothane concentration over the range 0.5% v/v halothane in nitrogen. Oxygen interferes with the measurement of halothane, but can be removed from a gas sample before analysis.

Electrodes↗

Pharmacological approach to the mechanisms of transcranial DC-stimulation-induced after-effects of human motor cortex excitability.

Weak transcranial direct current stimulation (tDCS) induces persisting excitability changes in the human motor cortex. These plastic excitability changes are selectively controlled by the polarity, duration and current strength of stimulation. To reveal the underlying mechanisms of direct current (DC)-induced neuroplasticity, we combined tDCS of the motor cortex with the application of Na(+)-channel-blocking carbamazepine (CBZ) and the N-methyl-D-aspartate (NMDA)-receptor antagonist dextromethorphan (DMO). Monitored by transcranial magnetic stimulation (TMS), motor cortical excitability changes of up to 40% were achieved in the drug-free condition. Increase of cortical excitability could be selected by anodal stimulation, and decrease by cathodal stimulation. Both types of excitability change lasted several minutes after cessation of current stimulation. DMO suppressed the post-stimulation effects of both anodal and cathodal DC stimulation, strongly suggesting the involvement of NMDA receptors in both types of DC-induced neuroplasticity. In contrast, CBZ selectively eliminated anodal effects. Since CBZ stabilizes the membrane potential voltage-dependently, the results reveal that after-effects of anodal tDCS require a depolarization of membrane potentials. Similar to the induction of established types of short- or long-term neuroplasticity, a combination of glutamatergic and membrane mechanisms is necessary to induce the after-effects of tDCS. On the basis of these results, we suggest that polarity-driven alterations of resting membrane potentials represent the crucial mechanisms of the DC-induced after-effects, leading to both an alteration of spontaneous discharge rates and to a change in NMDA-receptor activation.

Adult↗

Homeostatic-like plasticity of the primary motor hand area is impaired in focal hand dystonia.

The excitability of inhibitory circuits in patients with writer's cramp is reduced at multiple levels within the sensorimotor system, including the primary motor hand area (M1). Although this may play a major role in the pathophysiology of writer's cramp, it is still unclear what factors may cause the imbalance between inhibition and excitation to arise. One possibility is that homeostatic mechanisms that keep cortical excitability within a normal physiological range are impaired. In eight patients with writer's cramp and eight healthy age-matched controls, we combined low-frequency repetitive transcranial magnetic stimulation (rTMS) with transcranial direct current stimulation (TDCS) to probe regional homeostatic plasticity of the left M1. Confirming our previous study (Siebner et al., J Neurosci 2004; 24: 3379-85), 'facilitatory' preconditioning of the M1 with anodal TDCS enhanced the inhibitory effect of subsequent 1 Hz rTMS on corticospinal excitability. Conversely, 'inhibitory' preconditioning with cathodal TDCS reversed the after effect of 1 Hz rTMS, producing an increase in corticospinal excitability. The results were quite different in patients with writer's cramp. Following preconditioning with TDCS, 1 Hz rTMS induced no consistent changes in corticospinal excitability, indicating a loss of the normal 'homeostatic' response pattern. In addition, the normal inhibitory effect of preconditioning with cathodal TDCS was absent. The present data suggest that homeostatic mechanisms that stabilize excitability levels within a useful dynamic range are impaired in patients with writer's cramp. We propose that a faulty homeostatic response to acute increases in corticospinal excitability favours maladaptive motor plasticity. The role of homeostatic-like plasticity in the pathophysiology of task-specific dystonias warrants further study.

Adult↗

Site of initial excitation and current threshold as a function of electrode radius in heart muscle.

End-diastolic current thresholds have been measured in 13 open chested dogs as a function of electrode radius by stimulating the left ventricle with epicardial disc electrodes ranging in a radius from 0-3 mm to 9 mm. Thresholds for cathodal rectangular short stimuli as well as specifically for cathodal make stimulation, proved to be proportional to the electrode radius to the power 1-5. This relationship between radius and threshold can be explained theoretically, assuming that electrical stimulation results in a propagated depolarization front if a critical current density is reached somewhere in the myocardium. The current distribution measured over the electrode and the site of initial depolarization in the tissue are in accordance with this theoretical explanation.

Animals↗

Induction and the possible mechanism of ventricular tachycardia after catheter ablation with direct current shocks in dogs.

Cathodal DC shocks (150-J) were administered via the His bundle to 20 closed-chest dogs, and in a further three dogs 25-J cathodal shocks were given via the left ventricular endocardium. In 18 dogs, including three that underwent left ventricular ablation, Holter electrocardiograms were recorded from 1 to 7 days after ablation, and 4 weeks after ablation. There were frequent episodes of sustained ventricular tachycardia (VT) from the first few hours after ablation to 4 days after ablation in all dogs, but both the rate and the coupling interval of VT were variable. In five conscious dogs stimulated 1 day after ablation, it was difficult to induce and terminate VT repeatedly. There was a direct relationship between the paced cycle length and the interval of the last paced beat to the initiating VT beat in three out of four dogs. In the fourth dog there was an inverse relationship. There was no transient entrainment with ventricular burst pacing during VT in any of the four dogs tested. The effects of lidocaine (2-3 mg.kg-1), verapamil (0.2-0.4 mg.kg-1), and propranolol (0.2 mg.kg-1) on VT were tested within 2 days of ablation in 10 conscious dogs. In general, both lidocaine and verapamil terminated VT, and propranolol slowed VT. In conclusion, VT soon after ablation possibly results from triggered activity, although abnormal automaticity cannot be ruled out.

Animals↗

Analysis of a mouse alpha-globin gene mutation induced by ethylnitrosourea.

A DBA/2 mouse treated with ethylnitrosourea sired an offspring whose hemoglobin showed an extra band following starch gel electrophoresis. The variant hemoglobin migrated to a more cathodal position in starch gel. Isoelectric focusing indicated that chain 5 of the mutant hemoglobin migrated to a more cathodal position than the normal chain 5 from DBA/2 mice and that the other alpha-globin, chain 1, was not affected. On focusing gels the phenotype of the mutant allele, Hbay9, was expressed without dominance to normal chain 5, and Hbay9/Hbay9 homozygotes were fully viable in the laboratory. The molecular basis for the germinal mutation was investigated by analyzing the amino acid sequence of chain 5y9, the mutant form of alpha-chain 5. A single amino acid substitution (His leads to Leu) at position 89 was found in chain 5y9. We propose that ethylnitrosourea induced an A leads to T transversion in the histidine codon at position 89 (CAC leads to CTC). This mutation has apparently not been observed previously in humans, mice or other mammals, and its novel occurrence may be indicative of other unusual mutational events that do not ordinarily occur in the absence of specific mutagen exposure.

Amino Acid Sequence↗

Collection of gravitropic effectors from mucilage of electrotropically-stimulated roots of Zea mays L.

We placed agar blocks adjacent to tips of electrotropically stimulated primary roots of Zea mays. Blocks placed adjacent to the anode-side of the roots for 3 h induced significant curvature when subsequently placed asymmetrically on tips of vertically-oriented roots. Curvature was always toward the side of the root unto which the agar block was placed. Agar blocks not contacting roots and blocks placed adjacent to the cathode-side of electrotropically stimulated roots did not induce significant curvature when placed asymmetrically on tips of vertically-oriented roots. Atomic absorption spectrophotometry indicated that blocks adjacent to the anode-side of electrotropically-stimulated roots contained significantly more calcium than (1) blocks not contacting roots, and (2) blocks contacting the cathode-side of roots. These results demonstrate the presence of a gradient of endogenous Ca in mucilage of electrotropically-stimulated roots (i.e. roots undergoing gravitropic-like curvature).

Calcium↗