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The direction of neurite growth in a weak DC electric field depends on the substratum: contributions of adhesivity and net surface charge.

We investigated the influence of the growth surface on the direction of Xenopus spinal neurite growth in the presence of a dc electric field of physiological magnitude. The direction of galvanotropism was determined by the substratum; neurites grew toward the negative electrode (cathode) on untreated Falcon tissue culture plastic or on laminin substrata, which are negatively charged, but neurites growing on polylysine, which is positively charged, turned toward the positive electrode (anode). Growth was oriented randomly on all substrata without an electric field. We tested the hypothesis that the charge of the growth surface was responsible for reversed galvanotropism on polylysine by growing neurons on tissue culture dishes with different net surface charges. Although neurites grew cathodally on both Plastek substrata, the frequency of anodal turning was greater on dishes with a net positive charge (Plastek C) than on those with a net negative charge (Plastek M). The charge of the growth surface therefore influenced the frequency of anodal galvanotropism but a reversal in surface charge was insufficient to reverse galvanotropism completely, possibly because of differences in the relative magnitude of the substratum charge densities. The influence of substratum adhesion on galvanotropism was considered by growing neurites on a range of polylysine concentrations. Growth cone to substratum adhesivity was measured using a blasting assay. Adhesivity and the frequency of anodal turning were graded over the range of polylysine concentrations (0 = 0.1 < 1 < 10 = 100 microg/ml). The direction of neurite growth in an electric field is therefore influenced by both substratum charge and growth cone-to-substratum adhesivity. These data are consistent with the idea that spatial or temporal variation in the expression of adhesion molecules in embryos may interact with naturally occurring electric fields to enhance growth cone pathfinding.

Animals↗

Embryonic Xenopus neurites integrate and respond to simultaneous electrical and adhesive guidance cues.

Nerve cells detect and respond to multiple extrinsic guidance cues during development and regeneration using a motile growth cone. Navigational decisions may be required of the growth cone when different guidance cues are encountered simultaneously. We have tested the relative potencies of two opposing cues by presenting Xenopus spinal cord nerve cells growing on a micropatterned laminin culture substratum with an orthogonal DC electric field. Substrata composed of repeating 25-micron laminin tracks and spaces failed to influence the position of neuritogenesis from nerve cell soma. Once established, however, growth cone movement was constrained by laminin tracks such that neurites of 65% of cells were aligned after 5 h in vitro. Two hours after the application of a 100-140 mV/mm DC field the majority of cells remained aligned with the laminin tracks. Around 70% of Xenopus neurites normally orient cathodally on homogenous laminin substrata; therefore the galvanotropic response was impeded by prior exposure to a patterned laminin substrate. However, a proportion of aligned neurites did orient cathodally and evidence of a response to both directional cues was even found within the same cell. Video-enhanced contrast, differential interference contrast (VEC-DIC) microscopy was used to examine the detailed behavior of growth cones on micropatterned laminin substrata. The present study has demonstrated that growth cones can detect and integrate at least two morphogenetic guidance cues simultaneously. The strength of the galvanotropic response in Xenopus growth cones, however, was often insufficient to override established adhesive guidance in this model system.

Animals↗

The Comet-Tail (A(2)Pi(i)-X(2)Sigma(+)) System of (12)C(16)O(+): A Reinvestigation.

The emission spectrum of the comet-tail (A(2)Pi(i)-X(2)Sigma(+)) system of (12)C(16)O(+) produced in the cathode glow of a hollow-cathode discharge tube and photographed in the spectral region 3345-8500 Å at high resolution was reinvestigated. Detailed rotational analysis of 10 bands involving 5 vibrational levels v = 0 to 4 of each of the A and X states was carried out. Spin-splitting of the rotational levels was observed for the analyzed bands. The observed intensity distribution of the branches is found to be in good agreement with theoretical calculations. Molecular constants for both the A and X states were obtained from a global fit of the present data of the A-X system and the infrared and microwave data available from the literature for v = 0, 1, 2, 3, and 4 of state X, using effective Hamiltonian for the A(2)Pi state and matrix elements for both A and X states. The present work suggests that the A(2)Pi(i) state changes from Hund's case (a) at low J values to Hund's case (b) at high J values. Copyright 2000 Academic Press.

Journal Article↗

Introduction of DNA into chick embryos by in ovo electroporation.

Gene transfer by in ovo electroporation has been applied to the study of developmental biology, especially to central nervous system (CNS) development. Plasmids are injected into the neural tube of stage 10 chick embryos, and a 25-V 25-msec square pulse is applied five times. Since DNA moves toward the anode, the cathode side of the neural tube is transfected, and the cathode side is used as the control. Expression of translation product of the introduced DNA is observed 2 h after electroporation, peaks around 20 h after electroporation and then weakens. Expression is transient when plasmids are used as expression vectors, but they are very suitable for studying early developmental events (e.g., gene expression cascades or interactions). Misexpression of Pax-5 is shown as an example.

Animals↗

A new measuring device for non-invasive determination of oxygen partial pressure and oxygen conductance of the skin and other tissues.

About fifty percent of the oxygen consumption of the skin is supplied by diffusion through the surface. This portion of the skin oxygen supply becomes of high importance in case of arterial occlusion. The oxygen permeation coefficient (P) of the upper layers and the oxygen pressure field within the skin determine the diffusive oxygen uptake from the outside. To our knowledge, the permeation coefficient (P) until now was only estimated by indirect methods of little practicability (Baumberger et al., 1951; Eberhard et al., 1978). An oxygen partial pressure of the skin is conventionally measured by modified CLARK type electrodes. A disadvantage of this so-called transcutaneous electrode is its oxygen consumption and the fixed coupling of the consumption with the oxygen pressure to be determined. Therefore the measurement always induces a systematic error (the so-called stirring effect) which depends, among other factors, on the oxygen availability of the skin under the electrode. The new device combines a consumption-free oxygen partial pressure detector on the basis of luminescence quenching by oxygen with an independently working specific oxygen consumer realized by an active galvanic chain (silver-lead element). The chain permits setting any oxygen mass flow (mO2) in a certain range by varying the electrode current choosing different resistors within the electrical circuit. According to the diffusion law, the surface oxygen pressure (ePO2) being measured is a linear function of the oxygen flow (mO2) directed to the cathode: ePO2 identical to -(1/P).(mO2/A) + icPO2; A: area under the cathode. The intracutaneous oxygen partial pressure (icPO2) is a virtual quantity defined by the equation given. Only by using an active electrode different oxygen mass flows can be set and so the oxygen conductance of the upper skin layers can be assessed. First experiments on human skin in the gluteal region of an adult delivered an estimated value of the permeation coefficient (P): 2.2.10(-5) ml O2 (STPD)/(atm.s.cm2) at 42 degrees C skin surface temperature; the intracutaneous partial pressure obtained was 5.5 kPa (41 mmHg) (STPD: "standard temperature pressure dry" conditions of the gas). At 42 degrees C skin temperature no burning occurs. The determined O2-conductance is in the same range as estimated formerly (see above). The intracutaneous oxygen partial pressure determined seems to be a realistic value of the tissue at 42 degrees C. By in vitro measurements with technical membranes the new device and procedure was validated giving precise values of the oxygen conductance. Hence the new method may be used for calibration of the oxygen flux optode (Holst et al., 1993). The O2-permeation coefficient (P) could be an important parameter for evaluating dermatological applications (which attempt to increase P) in the treatment of local dermal oxygen deficiency. The intracutaneous oxygen partial pressure found is a reasonable value for a surface temperature of 42 degrees. Because of the independence of the O2-partial pressure determined and the O2-consumption the new device exhibits no stirring effect and it provides more insight into the limitations of conventional transcutaneous oxygen measurement.

Adult↗

Compensation of horizontal canal related activity in the medial vestibular nucleus following unilateral labyrinth ablation in the decerebrate gerbil. I. Type I neurons.

The spontaneous activity and dynamic responses to two frequencies (1.3 and 0.13 Hz) of sinusoidal angular horizontal head acceleration of type I neurons in the medial vestibular nucleus were recorded bilaterally in decerebrate Mongolian gerbils (Meriones unguiculatus) under three experimental conditions; normal labyrinth intact, acutely following unilateral labyrinthine lesion, and four to seven weeks following labyrinthine lesion. The mean spontaneous activity and number of detected type I neurons decreased immediately ipsilateral to the lesion but recovered significantly with time. In contrast, spontaneous activity on the contralateral side increased during compensation following hemilabyrinthectomy. The mean response gains at both frequencies of head oscillation were depressed bilaterally and asymmetrically acutely following the lesion such that the response gain of cells on the intact side exceeded that of the neurons recorded on the injured side. After compensation the number of detected type I neurons on the side ipsilateral to the injury increased but remained below normal levels. The mean gains remained depressed but became symmetric with compensation as a result of improvement in the response of ipsilateral neurons. The phase of responses were significantly advanced in the compensated animals. Although response gain is not fully restored, the linearity of the dynamic modulation in compensated animals is improved as evidenced by a continuous modulation of the increased spontaneous activity of neurons contralateral to the hemilabyrinthectomy. It is proposed that this effect is related to the concurrent improvement in the linearity of the horizontal vestibulo-ocular response. Electrical cathodal polarization of the vestibular nerve ipsilateral to the ablated labyrinth was utilized to investigate the relationship between recovery of spontaneous activity and dynamic function. Acutely following hemilabyrinthectomy, cathodal polarization restored activity in second-order type I neurons to near normal levels but their response gain to head rotation remained depressed. Similar galvanic stimulation in compensated animals also elevated ipsilateral spontaneous activity. As in the acute preparation, such stimulation did not modify the response gain or phase. Thus, the improvement in response of type I neurons in the compensated gerbil was not a direct consequence of restoration of spontaneous activity on the side of the injury.

Animals↗

A new hereditary variant of the PGM1 erythrocyte enzyme system determined by isoelectric focusing.

A new variant of the PGMa1 erythrocyte enzyme system not identical with the known variants of the system has been detected in the hemolyzed red blood cells of a healthy blood donor by isoelectric focusing. Using this technique the variant is represented by two bands, a strong and slow one more cathodically located than the a3 band and a weak one in the position of the a2 band. Using agarose thin layer or acetate foil electrophoresis the variant is represented only by a minimal cathodic broadening of the PGM1 1 band and therefore it is easily overlooked. Investigation of the propositus' family shows that the variant occurs combined with the common alleles PGMa1(1), PGMa1(2), and PGMa1(3) and that it has an autosomal dominant inheritance. Obviously the variant has a very low frequency.

Adult↗

Two subtypes of BfF by isoelectrofocusing: differential linkage to other HLA markers.

By isoelectrofocusing in agarose, the properdin factor allotype BfF could be split into two subtypes: BfFa with one major cathodic band and BfFb with the same cathodic band but in addition a major anodic band. By scanning, BfFaFb heterozygotes were distinguished from BfFbFb homozygotes by the stronger intensity of the anodic band in the latter. The two subtypes were segregated perfectly with HLA in 40 families and showed different association patterns with HLA markers. BfFa seemed to be linked to B35 while BfFb showed a strong linkage with all the components of the following haplotype: HLA-A29, Cw-, B44, BfFb, C4A3B1, DR7. The frequency of BfFa among BfFS heterozygotes was 41% (46/113) and that of BfFb 59% (67/113).

Alleles↗

PI*LBEI and PI*JHOU: two new alpha-1-antitrypsin alleles.

Two new variants of alpha-1-antitrypsin were found in a Chinese population by isoelectric focusing in polyacrylamide gel. They have been named Lbeijing (PI*LBEI) and Jhouyao (PI*JHOU) respectively. With the samples reduced and alkylated, both the variants were anodal to M1 and cathodal to L. Omitting the alkylation step, however, the relative mobilities of L and Jhouyao were altered. Under this condition, a characteristic of Jhouyao was revealed by isoelectric focusing in immobilized pH gradient 4.4-4.9, i.e., while its band 6 became anodal to L, band 4 was still cathodal to L.

Alleles↗

Glucose phosphate isomerase deficiency with hereditary hemolytic anemia in a Spanish family: clinical and familial studies.

A new case of glucose phosphate isomerase deficiency associated with cogenital nonspherocytic hemolytic anemia is described in a 12-year-old girl of Spanish origin. The parents exhibited erythrocyte glucose phosphate isomerase activity between 50 and 60% of normal. The enzyme of the propositus had normal Michaelis-Menten constants both for F-6-P and G-6-P, but abnormal pH optimum and decreased heat stability at 48 degrees C. On starch-gel electrophoresis the father's enzyme was normal but the mother's showed a cathodic migrating band in addition to the normal one. The enzyme from the propositus exhibited only one band with cathodal mobility of 116% of the main band found in normal subjects. It is postulated that the propositus is double heterozygous for two abnormal alleles, and the mother contributes a mutant allele with abnormal electrophoretic mobility and thermolability at 48 degrees C whereas the father contributes an allele without enzymatic activity.

Alleles↗

History of blood gas analysis. IV. Leland Clark's oxygen electrode.

The electrochemical reduction of oxygen was discovered by Heinrich Danneel and Walter Nernst in 1897. Polarography using dropping mercury was discovered accidentally by Jaroslav Heyrovsky in Prague in 1922. This method produced the first measured oxygen tension values in plasma and blood in the 1940s. Brink, Davies, and Bronk implanted platinum electrodes in tissue to study oxygen supply, or availability, from about 1940, but these bare electrodes became poisoned when immersed in blood. Leland Clark sealed a platinum cathode in glass and covered it first with cellophane; he then tested silastic and polyethylene membranes. In 1954 Clark conceived and constructed the first membrane-covered oxygen electrode having both the anode and cathode behind a nonconductive polyethylene membrane. The limited permeability of polyethylene to oxygen reduced depletion of oxygen from the sample, making possible quantitative measurements of oxygen tension in blood, solutions, or gases. This invention led to the introduction of modern blood gas apparatus.

Animals↗

Isoperoxidase spectra of single tulip cultivars and their parrot mutants.

The isoperoxidase spectrums of 2 single tulip cultivars and their parrot mutants were analyzed by disc electrophoresis in leaves and tepals i.e. petals plus sepals excised from within the bulbs. It was found that the anodal isoperoxidase pattern of parrot mutant tepals differs distinctly from tepals of their mother cultivars, but no differences were observed in cathodal isoperoxidases. Both the anodal and cathodal isoperoxidase spectra of parrot mutant leaves and leaves of their mother cultivars were similar.

Isoenzymes↗

Variation of alpha 1-antitrypsin glycoprotein microheterogeneity in hepatic postresuscitation disease.

Microheterogeneity of the glycoprotein alpha 1-antitrypsin has been investigated sequentially by high resolution isoelectric focusing in a child with the proteinase inhibitor MS phenotype after near-drowning. A band-splitting with additional cathodal fractions exhibited migration from the most cathodic to the anodic positions of the glycoprotein isoforms in the course of post-resuscitation disease. The pattern may reflect the time- and stage-dependent hypoxic and post-hypoxic effects on hepatocellular metabolic zonation.

Humans↗

Generation of unidirectionally propagating action potentials using a monopolar electrode cuff.

Unidirectionally propagating action potentials, which can be used to implement transmission failure on peripheral nerve through "collision block," have been generated electrically on cat myelinated peripheral nerve using a monopolar electrode cuff with the conductor positioned closest to the "arrest" end of the cuff. A single cathode located at least 5 mm from the arrest end resulted in unidirectional propagation with minimal current and charge injection. The range of stimulus current values that produced unidirectional propagation increased with increases in longitudinal asymmetry of cathode placement over the range of asymmetries tested (1.7:1 to 7:1). The stimulus current pulse that minimized charge injection was quasitrapezoidal in shape with a plateau pulse width of approximately 350 microseconds and an exponential trailing phase having a fall time (90%-10%) of approximately 600 microseconds These parameters were found to be independent of cuff geometry. Arrest efficiency was not degraded using a cuff of sufficient internal diameter to prevent nerve compression in chronic implantation. The critical current density within the extracellular space of the electrode cuff required to produce conduction failure at the arrest end was estimated to be 0.47 +/- 0.08 mA/mm2.

Action Potentials↗

Improved photomultiplier tube for positron emission tomography.

The paper describes an investigation in which it is shown that small positive voltage pulses applied to an external conductor placed against the photocathode of a photomultiplier tube can be used to switch the photocathode completely off for the duration of the pulses. This suggests that a photomultiplier tube with a multisegment photocathode can be constructed, the individual cathode segments of which can be switched off independently by means of such pulses. A theoretical explanation for the effect is provided with the aid of a simple circuit model for the photocathode. Analysis of the model also shows that it is possible to identify the particular cathode segment in which a photon is detected when a pulse is recorded at the phototube's anode. A phototube with these characteristics can have important implications for positron emission tomography, as it can provide improved spatial resolution, simultaneous multislice capability and the ability to eliminate distortion due to dead-time effects at high count rates.

Biomedical Engineering↗

Force recruitment during electrical nerve stimulation with multipolar intrafascicular electrodes.

Experimentally, during electrical nerve stimulation, the influence is examined of two intrafascicular anodes on the force recruitment with one intrafascicular cathode. It is found that the anodes suppress recruitment and that this effect is more pronounced when the distance of the anodes to the cathode is decreased, or when the anodal currents are increased. The measured recruitment curve patterns can be qualitatively explained by a nerve stimulation model that calculates theoretical recruitment curves for intrafascicular multi-electrode configurations. Discrepancies between the experimental and the theoretical recruitment curves are seen, but these can be understood by taking into account a non-uniform fibre distribution.

Animals↗

Transverse tripolar stimulation of peripheral nerve: a modelling study of spatial selectivity.

Various anode-cathode configurations in a nerve cuff are modelled to predict their spatial selectivity characteristics for functional nerve stimulation. A 3D volume conductor model of a monofascicular nerve is used for the computation of stimulation-induced field potentials, whereas a cable model of myelinated nerve fibre is used for the calculation of the excitation thresholds of fibres. As well as the usual configurations (monopole, bipole, longitudinal tripole, 'steering' anode), a transverse tripolar configuration (central cathode) is examined. It is found that the transverse tripole is the only configuration giving convex recruitment contours and therefore maximises activation selectivity for a small (cylindrical) bundle of fibres in the periphery of a monofascicular nerve trunk. As the electrode configuration is changed to achieve greater selectivity, the threshold current increases. Therefore threshold currents for fibre excitation with a transverse tripole are relatively high. Inverse recruitment is less extreme than for the other configurations. The influences of several geometrical parameters and model conductivities of the transverse tripole on selectivity and threshold current are analysed. In chronic implantation, when electrodes are encapsulated by a layer of fibrous tissue, threshold currents are low, whereas the shape of the recruitment contours in transverse tripolar stimulation does not change.

Animals↗

Discrimination of liver cirrhosis from chronic hepatitis by analysis of serum cholinesterase isozymes using affinity electrophoresis with concanavalin A or wheat germ agglutinin.

Isozymic alteration of serum cholinesterase (ChE) was investigated in patients with chronic liver diseases using affinity electrophoresis with concanavalin A (Con A) or wheat germ agglutinin (WGA). On Con A-containing agarose gel electrophoresis, three bands with enzyme activity (named bands I to III, from the anodic side to the cathodic) were observed in sera of normal controls. Disappearance of band II was observed in 50% (15/30) of cirrhotic patients, but only one of 20 patients with chronic hepatitis lacked band II of the serum ChE isozymes. Meanwhile, WGA-containing agarose gel electrophoresis revealed that normal controls had four ChE isozymes (named bands I to IV from the anodic side to the cathodic). These four isozymes were also observed in patients with chronic hepatitis. However approximately 67% (20/30) of cirrhotic patients lacked band II of ChE isozymes. When these two affinity electrophoreses were used in combination, 22 (73%) of 30 cirrhotic patients had isozymic alteration of their serum ChE on either Con A-containing or WGA-containing agarose gel electrophoresis, or both. Thus, affinity electrophoreses with Con A and WGA seemed to be useful methods in differentiating liver cirrhosis from chronic hepatitis.

Cholinesterases↗