Evidence that the Loss of the Voltage-Dependent Mg2+ Block at the N-Methyl-D-Aspartate Receptor Underlies Receptor Activation During Inhibition of Neuronal Metabolism

Both phosphointermediate- and vacuolar-type (P- and V-type, respectively) ATPase activities found in cholinergic synaptic vesicles isolated from electric organ are immunoprecipitated by a monoclonal antibody to the SV2 epitope characteristic of synaptic vesicles. The two activities can be distinguished by assay in the absence and presence of vanadate, an inhibitor of the P-type ATPase. Each ATPase has two overlapping activity maxima between pH 5.5 and 9.5 and is inhibited by fluoride and fluorescein isothiocyanate. The P-type ATPase hydrolyzes ATP and dATP best among common nucleotides, and activity is supported well by Mg2+, Mn2+, or Co2+ but not by Ca2+, Cd2+, or Zn2+. It is stimulated by hyposmotic lysis, detergent solubilization, and some mitochondrial uncouplers. Kinetic analysis revealed two Michaelis constants for MgATP of 28 microM and 3.1 mM, and the native enzyme is proposed to be a dimer of 110-kDa subunits. The V-type ATPase hydrolyzes all common nucleoside triphosphates, and Mg2+, Ca2+, Cd2+, Mn2+, and Zn2+ all support activity effectively. Active transport of acetylcholine (ACh) also is supported by various nucleoside triphosphates in the presence of Ca2+ or Mg2+, and the Km for MgATP is 170 microM. The V-type ATPase is stimulated by mitochondrial uncouplers, but only at concentrations significantly above those required to inhibit ACh active uptake. Kinetic analysis of the V-type ATPase revealed two Michaelis constants for MgATP of approximately 26 microM and 2.0 mM. The V-type ATPase and ACh active transport were inhibited by 84 and 160 pmol of bafilomycin A1/mg of vesicle protein, respectively, from which it is estimated that only one or two V-type ATPase proton pumps are present per synaptic vesicle. The presence of presumably contaminating Na+,K(+)-ATPase in the synaptic vesicle preparation is demonstrated.     

Development of the jamming avoidance response and its morphological correlates in the gymnotiform electric fish,Eigenmannia

The electric fish, Eigenmannia, will smoothly shift the frequency of its electric organ discharge away from an interfering electric signal. This shift in frequency is called the jamming avoidance response (JAR). In this article, we analyze the behavioral development of the JAR and the anatomical development of structures critical for the performance of the JAR. The JAR first appears when juvenile Eigenmannia are approximately 1 month old, at a total length of 13–18 mm. We have found that the establishment of much of the sensory periphery and of central connections precedes the onset of the JAR. We describe three aspects of the behavioral development of the JAR: (a) the onset and development of the behavior is closely correlated with size, not age; (b) the magnitude (in Hz) of the JAR increases with size until the juveniles display values within the adult range (10–20 Hz) at a total length of 25–30 mm; and (3) the JAR does not require prior experience or exposure to electrical signals. Raised in total electrical isolation from the egg stage, animals tested at a total length of 25 mm performed a correct JAR when first exposed to the stimulus. We examine the development of anatomical areas important for the performance of the JAR: the peripheral electrosensory system (mechano- and electroreceptors and peripheral nerves); and central electrosensory pathways and nuclei [the electrosensory lateral line lobe (ELL), the lateral lemniscus, the torus semicircularis, and the pacemaker nucleus]. The first recognizable structures in the developing electrosensory system are the peripheral neurites of the anterior lateral line nerve. The afferent nerves are established by day 2, which is prior to the formation of receptors in the epidermis. Thus, the neurites wait for their targets. This sequence of events suggests that receptor formation may be induced by innervation of primordial cells within the epidermis. Mechanoreceptors are first formed between day 3 and 4, while electroreceptors are first formed on day 7. Electroreceptor multiplication is observed for the first time at an age of 25 days and correlates with the onset of the JAR. The somata of the anterior lateral line nerve ganglion project afferents out to peripheral electroreceptors and also send axons centrally into the ELL. The first electroreceptive axons invade the ELL by day 6, and presumably a rough somatotopic organization and segmentation within the ELL may arise as early as day 7. Axonal projections from the ELL to the torus develop after day 18. Within the torus semicircularis, giant cells are necessary for the performance of the JAR. Giant cell numbers increase exponentially during development and the onset of the JAR coincides with a minimum of at least 150 giant cells and the attainment of a total length of at least 15 mm and at least 150 giant cells. Pacemaker and relay cells comprise the adult Eigenmannia pacemaker nucleus. The growth and differentiation of these cell types also correlates with the onset of the JAR in developing animals. We describe a gradual improvement of sensory abilities, as opposed to an explosive onset of the mature JAR. We further suggest that this may be a rule common in most developing behavioral systems. © 1992 John Wiley & Sons, Inc.     

Determination of electric current distributions in animals and humans exposed to a uniform 60-Hz high-intensity electric field

The thermographic method for determining specific absorption rate (SAR) in animals and models of tissues or bodies exposed to electromagnetic fields was applied to the problem of quantifying the current distribution in homogeneous bodies of arbitrary shape exposed to 60-Hz electric fields. The 60-Hz field exposures were simulated by exposing scale models of high electrical conductivity to 57.3-MHz VHF fields of high strength in a large 3.66 × 3.66 × 2.44-m TE₁₀₁ mode resonant cavity. After exposure periods of 2–30 s, the models were quickly disassembled so that the temperature distribution (maximum value up to 7 °C) along internal cross-sectional planes of the model could be recorded thermographically. The SAR, W′, calculated from the temperature changes at any point in the scale model was used to determine the SAR, W, for a full-scale model exposed to a 60-Hz electric field of the same strength by the relation W = (60/ f² · (σ′/σ) · W′ where f′ is the model exposure frequency, σ′ is the conductivity of the scale model at the VHF exposure frequency, and σ is the conductivity of the full-scale subject at 60 Hz. The SAR was used to calculate either the electric field strength or the current density for the full-scale subject. The models were used to simulate the exposure of the full-scale subject located either in free space or in contact with a conducting ground plane. Measurements made on a number of spheroidal models with axial ratios from 1 to 10 and conductivity from 1 to 10 s/m agreed well with theoretical predictions. Maximum current densities of 200 nA/cm² predicted in the ankles of man models and 50 nA/cm² predicted in the legs of pig models exposed to 60-Hz fields at 1kV/m agreed well with independent measurements on full-scale models.     

The Synthesis, Storage, and Release of Propionylcholine by the Electric Organ of Torpedo marmorata

Abstract: Little is known about the specificity of the mechanisms involved in the synthesis and release of acetylcholine for the acetyl moiety. To test this, blocks of tissue from the electric organ of Torpedo were incubated with either [1-¹⁴C]acetate or [1-¹⁴C]propionate, and the synthesis, storage, and release of [1-¹⁴C]acetylcholine and [¹⁴C]propionylcholine were compared. To obtain equivalent amounts of the two labeled choline esters, a 50-fold higher concentration of propionate than of acetate was needed. Following subcellular fractionation, similar proportions of [¹⁴C]acetylcholine and [¹⁴C]propionylcholine were recovered with synaptosomes and with synaptic vesicles. Furthermore, both labeled choline esters were protected to a similar extent from degradation during homogenization of tissue in physiological medium, indicating that the two choline esters were equally well incorporated into synaptic vesicles. Yet depolarization of tissue blocks by 50 m M KCI released much less [¹⁴C]propionylcholinc than [¹⁴C]acetylcholine. During field stimulation of the tissue blocks, the difference between the releasibility of the two choline esters was less marked, but acetylcholine was still released in preference to propionylcholine. Evidence for specificity of the release mechanism was also obtained when the release of the two choline esters in response to field stimulation was compared in tissue blocks preincubated with both [³H]choline and [¹⁴C]propionate.     

Effect of aliphatic alcohols on the conformation of poly (l-ornithine hydrobromide) as studied by square-pulse and reversing-pulse electric birefringence

The electric birefringence and circular dichroism spectra of poly(l-ornithine hydrobromide) have been measured in ethanol/water, 2-propanol/water and tertiary butyl alcohol/water mixtures of various compositions. This charged polypeptide underwent a transition from the coil conformation to the helical conformation at high alcohol content in every case tested. Anomalous birefringence signals, indicative of a field-induced helix-to-coil transition. were observed at high electric fields only in the case of ethanol/water mixtures. The reversing-pulse electric birefringence of this polypeptide has been studied in ethanol/water mixtures and in neutral aqueous solution. Upon rapid reversal of the pulse field, no transient could be observed. This confirms that the electric-field orientation of poly(l-ornithine hydrobromide) results predominantly from the contribution of the counterion-induced dipole moment, regardless of its molecular conformations. It is very probable that the backbone permanent dipole moment of the helical conformation is largely suppressed by the counterion-induced dipole moment in the ionized form.     

A pulsed NMR study of lipids,bound water and sodium ions in macroscopically-oriented lecithin/water lyotropic liquid crystal model membrane systems

The temperature and orientation dependence of pulsed NMR ‘free induction decay’ signals have been studied in detail for lipid bilayers macroscopically-oriented between glass slides. Results for the lipid molecules (¹H, ³¹P), bound water (²H₂O) and ions dissolved in the aqueous phase (²³Na) are presented. Bilayers of egg-lecithin, dimyristoyl lecithin and potassium oleate have been investigated. In the liquid crystal phase all the signals, including those from bound water and ions exhibit a |3 cos²? ? 1| dependence on orientation of the bilayer normal to the magnetic field. In the case of DML samples, some orientation dependence of both ¹H and ²H signals persists in the gel phase, indicating that the lipid molecules retain a degree of reorientational freedom about their long axes in this phase. At the gel-liquid crystal transition the ²H quadrupole spittings undergo a discontinuous change. Results are interpreted in terms of a model in which water molecules are bound to individual lipid head groups and reorient with them, while sodium ions are located in the aqueous channel between bilayers.     

Studies on the possible biological effects of 50 Hz electric and/or magnetic fields: Evaluation of some glycolytic enzymes,glycolytic flux,energy and oxido-reductive potentials in human erythrocytes exposed in vitro to power frequency fields

An attempt has been made to understand whether 50 Hz electric and magnetic fields (EMFs) are involved in producing bioeffects by exposing human erythrocytes in vitro. The study evaluated some key glycolytic enzymes, glucose consumption, lactate production, energy charge, 2,3-diphosphoglycerate, and reduced glutathione levels. all of which are biochemical parameters significant to erythrocyte function. Cells exposed to individual or superimposed EMFs have not shown any significant difference compared with the controls. © 1993 Wiley-Liss. Inc.     

Kinetic,isotherm and thermodynamic investigations on adsorption of trace elements and pigments from soybean oil using high voltage electric field-assisted bleaching: A comparative study

In this study, the amounts of trace elements (Fe (II) and Cu (II)) and pigments (chlorophyll and carotenoid) in soybean oil were evaluated under high voltage electric field (HVEF) bleaching method at different voltage (10 and 20 kV), temperature (35–65 °C), time (0−30 min) and clay percentage (0.5–2 %) and then were compared to the industrial bleaching method (IBM). The kinetic data of ions and pigments adsorbed on activated bentonite clay under IBM and HVEF at two voltages of 10 and 20 kV followed the mechanism of the pseudo-first-order model (PFOM). The carotenoid and chlorophyll equilibrium data followed a Freundlich isotherm type model, which demonstrated multilayer adsorption under HVEF. The thermodynamic parameters (ΔG°, ΔH°, and ΔS°) displayed that the adsorption of trace metal ions and pigments on bentonite clay under IBM and HVEF were feasible, endothermic and spontaneous between 35 and 65 °C. The results indicated that the HVEF, especially at higher voltage, has a high remarkable capability to remove metal ions and pigments from soybean oil than the IBM. The highest removal capacity for metal and pigments of soybean oil bleaching were obtained in the order of HVEF-20 kV > HVEF-10 kV > IBM.     

Technoecological analysis of energy carriers for long‐haul transportation

Long‐haul transportation demand is predicted to increase in the future, resulting in higher carbon dioxide emissions. Different drivetrain technologies, such as hybrid or battery electric vehicles, electrified roads, liquefied natural gas and hydrogen, might offer solutions to this problem. To assess their ecological and economic impact, these concepts were simulated including a weight and cost model to estimate the total cost of ownership. An evolutionary algorithm optimizes each vehicle to find a concept specific optimal solution. A model calculates the minimum investment in infrastructure required to meet the energy demand for each concept. A well‐to‐wheel analysis takes into account upstream and on‐road carbon dioxide emissions, to compare fully electric vehicles with conventional combustion engines. Investment in new infrastructure is the biggest drawback of electrified road concepts, although they offer low CO₂ emissions. The diesel hybrid is the best compromise between carbon reduction and costs.