EEG Effects of Therapeutic Electric Stimulation of the Human Brain in the Posttraumatic Unconscious State

The effects of therapeutic SCS and transcranial electric stimulation on the functional activity of the brain in seven patients in the posttraumatic unconscious state were compared. The therapeutic transcranial stimulation was shown to exert a positive effect on the EEG and the characteristics of its spatial–temporal organization in most cases, which corresponds with positive shifts in the mental and motor sphere. The phasic character of changes in the bioelectric activity reflecting the sequence of involvement of the cerebral structures (primary activation of the deep frontal lobe divisions, activation of the subcortical-diencephalic structures, activation of the cortex) in the developing adaptive reactions, which helps understand their neurophysiological mechanisms, was described. It was established that the formation of the foci of persistent pathological activity with dominant characteristics in the brain is one of the possible complications of both SCS and transcranial electric stimulation. The mechanisms of their neurogenesis and the electrographic equivalents require further study. The data may serve as the basis for further clinical study of the influence of transcranial electric stimulation on the recovery of patients in postcomatose unconscious states.     

Changes in the configuration of conditioned reactions of visual cortex neurons during changes in reinforcement parameters]

In order to study the influences of controlled changes of defensive integration on the activity of visual cortical units their responses to a conditioned light flash and electric cutaneous stimulation with a 600 msec interval between them were recorded in experiments on alert rabbits. It has been shown that in a third of the neurones the types of reaction to light flashes and electric stimuli coincide. The changes in parameters of the reinforcing shock led to a changed response of most cells to the conditioned photic stimulus and electric stimulation. The changes may have affected units which produce any activation phase, including cells with activity characteristic of detectory ("simple" and "complex") visual neurones. The data obtained suggest that the special function of the visual cortex is used in different ways in systemic mechanisms of conditioned and unconditioned defensive acts and that the integrated system of a behavioral act exerts control both on the use of the unit in a certain systemic process and on its receptive field.     

Induction of chemiluminescence of peritoneal exudate macrophages after exposure to an electric field

The effect of the external high voltage electric field pulses on the suspension of rat peritoneal phagocytes has been investigated using chemiluminescent and turbodimetric methods. Single electric field pulses were found to activate macrophages, which was accompanied by a "flash" of chemiluminescence. Subthreshold electric field strength up to 0.8 kV/cm failed to alter macrophage activity. Maximum activation was observed at 2.2 kV/cm; with higher electric field intensity the effect diminished to zero. Drastic changes in light-scattering suspension properties at high electric field intensity indicate irreversible alterations of the barrier function of phagocyte membranes.     

Monoamine oxidase activity in rabbit brain nuclei in experimental acute emotional stress

Activity of monoamine oxidase in the neurons of catecholamine-synthesizing nuclei of hypothalamic and brainstem of rabbits with different resistance of cardiovascular functions to emotional stress was studied by the Glenner et al. method. An acute experimental emotional stress was induced by non-periodic electric stimulation of the hypothalamic ventromedial nucleus and the skin in immobilized adult chinchilla rabbits. It was found that monoamine oxidase activity in the neurons of groups A5, A6, A7 and A14 under acute experimental emotional stress increased in rabbits "resistant"to stress and remained unchanged in rabbits "predisposed" to stress. The activity of monoamine oxidase in the neurons of group A12 remained unchanged in the rabbits "resistant"to stress and decreased in the rabbit "predisposed" to stress.     

Electrosensory modulation of escape responses

Once initiated, rapid escape responses of teleost fishes are thought to be completed without additional sensory modification. This suggests that the motor program for a particular response is selected for by the constellation of sensory cues existing at the time of the releasing stimulus. This paper presents initial evidence that a highly specialized, phylogenetically recent electrosensory system is integrated with a primitive motor system and allows an animal to continuously monitor its environment for producing accurate escape behaviors.Behavioral testing for directed startle responses in a Y-maze demonstrates that when presented immediately before an acoustic startle stimulus, electric fish (Eigenmannia virescens), direct their response away from the cue (a transient shorting of their electric field). Thus, electrosensory cues as brief as 100 ms provide directional information to the escape motor network.In electric fish that are curarized to facilitate intracellular recording, the normal electric organ discharge is attenuated. When an electronically generated replacement field of the same frequency and amplitude as the fish's normal signal is shorted, a fast-rising, 7 ms latency post-synaptic potential is evoked from the Mauthner cell. Similar PSPs are generated by turning the replacement stimulus on and off. In some recordings, removing the S1 replacement field elicits a rebound of other afferent activity to the Mauthner cell; replacing the field suppresses this activity.Abbreviations EHP extrinsic hyperpolarizing potential - EOD electric organ discharge - JAR jaming avoidance response - LED light emitting diode - PSP postsynaptic potential     

Distinct mechanisms of modulation in a neuronal oscillator generate different social signals in the electric fishHypopomus

Summary The medullary pacemaker nucleus of the gymnotiform electric fish,Hypopomus, is a relatively simple neuronal oscillator which contains pacemaker cells and relay cells. The pacemaker cells generate a regular discharge cycle and drive the relay cells which trigger pulse-like electric organ discharges (EODs). The diencephalic prepacemaker nucleus (PPN) projects to the pacemaker nucleus and modulates its activity to generate a variety of specific discharge patterns which serve as communicatory signals (Figs. 2 and 3).While inducing such signals by microiontophoresis of L-glutamate to the region of the PPN (Fig. 4) of curarized animals, we monitored the activity of neurons in the pacemaker nucleus intracellularly. We found that pacemaker cells and relay cells were affected differently in a manner specific to the type of EOD modulation (Figs. 5–10). The normal sequence of pacemaker cell and relay cell firing was maintained during gradual rises and falls in discharge rate. Both types of cells ceased to fire during interruptions following a decline in discharge rate. During sudden interruptions, however, relay cells were steadily depolarized, while pacemaker cells continued to fire regularly. Short and rapid barrages of EODs, called chirps, were generated through direct and synchronous activation of the relay cells whose action potentials invaded pacemaker cells antidromically and interfered with their otherwise regular firing pattern.Abbreviations EOD electric organ discharge - HRP horseradish peroxidase - NMDA N-Methyl-D-Aspartate - PPN prepacemaker nucleus     

The direct influence of electromagnetic fields on nerve- and muscle cells of man within the frequency range of 1 Hz to 30 MHz

Summary By using several biophysical approximations and considering man as free space model limiting order-of-magnitude values for external electric and magnetic field strengths which may be hazardous for human beings were calculated. Danger may occur by excitation processes below 30 kHz for field strengths exceeding these limiting values; for frequencies larger than 30 kHz, thermal effects are predominant before excitation occurs. The external electric field strength necessary for causing action potentials in the central nervous system exceeds by far the corona forming level. But excitation is possible by strong alternating magnetic fields.Furthermore, by comparing the electrically and magnetically induced currents with the naturally flowing currents in man caused by the brain's and heart's electrical activity, a lower boundary-line was estimated. Regarding electric or magnetic field strengths undercutting this boundary-line, direct effects on the central nervous system may be excluded. Other mechanisms should be responsible for demonstrated biological effects.     

EMG activity of pigeon oesophagus in vivo

At rest, the pigeon cervical oesophagus, which is entirely smooth muscle, shows electric activity. This activity consists of bursts of spikes with frequency increasing in the oral-aboral direction. The bursts are un-phase locked, and there are no slow waves (E.C.A.). The surgical transection of the oesophageal muscular wall does not affect the electric activity even in a disconnected segment. After asphyxia electric activity persists, whereas the aboral gradient of frequency disappears. Therefore, the electric activity is thought to be myogenic in origin, and the frequency gradient nervous in origin. Atropine and neostigmine administration suggests that the cholinergic system modulates the electric activity, but it is not involved in the control of the frequency gradient. On the contrary, hexamethonium administration, by abolishing this gradient, lends support to the idea of a postganglionic atropine-resistant neuronal system responsible for the gradient.     

Reversible inhibition of acetylcholinesterases of different origins by quaternary phosphonium compounds

Studies have been made on the reversible inhibition of acetylcholinesterase activity from the erythrocytes of man, horse and camel, the electric organ of the skate Torpedo marmorata and eel Electrophorus electricus, the venom of the snakes Naja naja and Vipera lebetina, the brain of the pigeon Columba livia by tetraphenyl-, triphenylalkyl- and tributyrylalkyl-phosphonium salts. The investigated phosphonium inhibitors exhibit an evident specificity in their action: they were more effective in inhibiting the acetylcholinesterase from human erythrocytes than that from the erythrocytes of horse and camel. These salts were more effective with respect to the acetylcholinesterase activity of the electric organ of the skate than that of the electric organ of the eel. Acetylcholinesterases from the venom of the snakes exhibited practically identical sensitivity to all the phosphonium compounds investigated. The present work is the first attempt to use quaternary phosphonium salts (the so-called "hydrophobic ions") in comparative enzymological investigation.     

A study of the parameters of electric stimulation on the dorsal hippocampus for the "after discharge", carried out with the aid of a current-operated telestimulator, in the rabbit supplied with permanent electrodes

Our purpose in this work has been to carry out experiment of electric stimulation of the dorsal hippocampus, by measuring exactly the quantity of stimulating current, expressed in mA, in order to ascertain the actual quantities of electric charges, expressed in microcoulombs and conveyed to the examined structure and able to cause "an after discharge". the optimum stimulation frequency involving supply of the lowest quantity of electric charges and such frequency is more or less equal to the frequency of discharge of the paroxysmal bioelectric activity (about 20 c/sec). The optimum length of the train of stimulation is very short, but non shorter than 1 sec; consequently the length of the train of stimuli of 2, 5 sec, corresponding to a current intensity inferior to 1 mA and to a sufficiently low quantity fo charges, is to be regarded as the optimum length. The optimum length of the single stimulus is very short too, but it cannot be too short, since, also in this case, the intensity of current necessary to reach the threshold for the "after discharge" would reach such values as to cause lesions in the stimulated structure; consequently the length of the single stimulus corresponding to 0, 4 msec is to be considered as to optimum length.     

Disorder of electromechanical coupling in the cells of the myocardium in protracted crush syndrome

Experiments on papillary muscles of normal (control) rabbits and of those with the compression syndrome (CS) were made to explore the action of the control and "syndromic" blood plasma on electric and contractile activity of the myocardium. Isometric contractions of myocardial preparations were recorded at varying stimulation frequencies (0.1-2 Hz). Intracellular rest potentials (RP) and action potentials (AP) were led away with the aid of glass microelectrodes filled with 2.5 M KCl. The replacement of Tyrode solution by the control plasma raised the amplitude of papillary muscle contractions, that being greater as regards the muscles from rabbits with the CS. The "syndromic" plasma (diluted by Tyrode solution in a 1:1 ratio) markedly inhibited the amplitude of contractions of papillary muscles from both the control rabbits and animals with the CS. Reduction of the contractions induced by the "syndromic" plasma seen in all the preparations was followed by two patterns of changes in electrical activity of myocardial fibers. In one pattern, the RP, the amplitude and duration of the AP declined. In the other, on the contrary, the changes were reduced to a greater AP duration. The conclusion is made about the absence of a direct relationship between the decrease in myocardial contractility and changes in intracellular potentials induced by the "syndromic" plasma. It is suggested that the "syndromic" plasma deranges the process of stimulation and contraction coupling in heart papillary muscles.     

The release of adenosine at the electric organ ofTorpedo. A study using a continuous chemiluminescent method

Acetylcholine and ATP are costored and coreleased during synaptic activity at the electric organ ofTorpedo. It has been suggested that released ATP is converted to adenosine at the synaptic cleft, and in turn this nucleoside would depress the evoked release of acetylcholine. In the present communication we have used a chemiluminescent reaction that let us to monitor continuously the presence of adenosine in this preparation. The chemiluminescent reaction is based on the conversion of adenosine into uric acid and H₂O₂ by adenosine deaminase, nucleoside phosphorylase, and xanthine oxidase enzymes. The hydrogen peroxide has been detected by peroxidase-luminol mixture. The reaction has a sensitivity on the picomol range and discerned between Adenosine, AMP, ADP, and ATP. We have developed this technique in the hope of understanding whether adenosine is released during synaptic activity or it comes from the released ATP. We have studied the release or formation of adenosine in fragments of the electric organ and in isolated cholinergic nerve terminals obtained from it. In both conditions we have followed the effect of potassium stimulation upon the detection of adenosine. Potassium stimulation increased the extracellular adenosine either in slices or the synaptosomal fraction ofTorpedo electric organ. The presence of , -methylene ADP, an inhibitor of 5-nucleotidase, inhibits the detection of adenosine, suggesting that extracellular adenosine is a consequence of ectocellular dephosphorylation of released ATP.     

Changing the Direction and Orientation of Electric Field During Electric Pulses Application Improves Plasmid Gene Transfer in vitro

Gene electrotransfer is a physical method used to deliver genes into the cells by application of short and intense electric pulses, which cause destabilization of cell membrane, making it permeable to small molecules and allows transfer of large molecules such as DNA. It represents an alternative to viral vectors, due to its safety, efficacy and ease of application. For gene electrotransfer different electric pulse protocols are used in order to achieve maximum gene transfection, one of them is changing the electric field direction and orientation during the pulse delivery. Changing electric field direction and orientation increase the membrane area competent for DNA entry into the cell. In this video, we demonstrate the difference in gene electrotransfer efficacy when all pulses are delivered in the same direction and when pulses are delivered by changing alternatively the electric field direction and orientation. For this purpose tip with integrated electrodes and high-voltage prototype generator, which allows changing of electric field in different directions during electric pulse application, were used. Gene electrotransfer efficacy is determined 24h after pulse application as the number of cells expressing green fluorescent protein divided with the number of all cells. The results show that gene transfection is increased when the electric field orientation during electric pulse delivery is changed.Download video file.^{(27M, mov)}     

Interrelationship between the neocortex and limbic system in emotional stress and epileptiform syndromes

The paper describes bioelectrical correlates of negative emotional states on three models: 1) states caused by local, chronic electric stimulation of negative emotiogenic zones (in cats, rabbits); 2) by a neurogenic stress in rats resulting from a conflict of afferent excitations; 3) by an experimental syndrome--"expectation stress". The data attest that hypersynchronization of bioelectrical activity in limbic and cortical structures is a sufficiently significant correlate of an emotional stress. It is assumed that hypersynchronization reflects one of the general mechanisms of involvement of brain structures in a state of neurogenic stress and its manifestation.     

Theoretical analysis of pre-receptor image conditioning in weakly electric fish

Electroreceptive fish detect nearby objects by processing the information contained in the pattern of electric currents through the skin. The distribution of local transepidermal voltage or current density on the sensory surface of the fish's skin is the electric image of the surrounding environment. This article reports a model study of the quantitative effect of the conductance of the internal tissues and the skin on electric image generation in Gnathonemus petersii (Günther 1862). Using realistic modelling, we calculated the electric image of a metal object on a simulated fish having different combinations of internal tissues and skin conductances. An object perturbs an electric field as if it were a distribution of electric sources. The equivalent distribution of electric sources is referred to as an object's imprimence. The high conductivity of the fish body lowers the load resistance of a given object's imprimence, increasing the electric image. It also funnels the current generated by the electric organ in such a way that the field and the imprimence of objects in the vicinity of the rostral electric fovea are enhanced. Regarding skin conductance, our results show that the actual value is in the optimal range for transcutaneous voltage modulation by nearby objects. This result suggests that "voltage" is the answer to the long-standing question as to whether current or voltage is the effective stimulus for electroreceptors. Our analysis shows that the fish body should be conceived as an object that interacts with nearby objects, conditioning the electric image. The concept of imprimence can be extended to other sensory systems, facilitating the identification of features common to different perceptual systems.     

Rearrangements of the Total Electric Activity of the Cerebral Cortex and Subcortical Structures in Experimental Hypoxia

Rearrangements of the parameters of the electric activity of the cortical and subcortical regions were studied at different stages of experimental hypoxia (exposure to oxygen–nitrogen mixtures with an oxygen content of 7.5–8.0%) in chronic experiments of rabbits with the use of electrodes implanted into the brain. Acute hypoxia was shown to cause characteristic changes in electrical activity in the cortex, reticular formation, caudate nuclei, and hippocampus. The sequence of these changes may be divided into the following stages: (1) a short-term activation of all cerebral structures during the first 15–30 s and a shift of the frequency spectrum towards higher frequencies ( and waves), (2) a decrease in and waves and an increase in activity during the next 2–4 min, and (3) a gradual shift towards slow waves (the rhythm) and paroxysmal episodes in some animals. These shifts first appear in the frontal cortical regions and the reticular formation and then in the caudate nuclei and hippocampus. The degree of the changes in electric activity is correlated with the decrease in oxygen tension in the arterial blood.     

Electric field effect on the zigzag (6,0) single-wall BC2N nanotube for use in nano-electronic circuits

We have analyzed the effect of external electric field on the zigzag (6,0) single-wall BC₂N nanotube using density functional theory calculations. Analysis of the structural parameters indicates that the nanotube is resistant against the external electric field strengths. Analysis of the electronic structure of the nanotube indicates that the applied parallel electric field strengths have a much stronger interaction with the nanotube with respect to the transverse electric field strengths and the nanotube is easier to modulate by the applied parallel electric field. Our results show that the properties of the nanotube can be controlled by the proper external electric field for use in nano-electronic circuits.

Creatine phosphokinase: isoenzymes in Torpedo marmorata

Creatine phosphokinase (ATP: creatine N-phosphotransferase, EC 2.7.3.2) is the major constituent of the "low-salt-soluble" proteins of the electric organ from Torpedo marmorata. The denatured subunits of the enzyme have an apparent Mr of 43 000 and isoelectric points ranging between pH 6.2 and pH 6.5. Identical properties are found for the creatine phosphokinase from Torpedo muscle tissue. Anti-(electric organ creatine phosphokinase) antibodies are specific for the muscle-type enzyme and do not cross-react with enzymes present in Torpedo brain and electric lobe tissue. Biochemical and immunochemical properties of the enzyme associated with acetylcholine-receptor-enriched membranes show that this enzyme is as the "low-salt-soluble" electric organ enzyme of the muscle-specific type. In vitro translation of electric organ poly(A)-rich mRNA in a reticulocyte lysate reveals the abundance of mRNA specific for muscle creatine phosphokinase. During embryonic development of the electrocyte a continuous increase of translatable amounts of this mRNA is observed. No brain-type polypeptides are synthesized. The subunits of the brain-specific enzyme differ in molecular mass (Mr approximately equal to 42000) and isoelectric properties (pI approximately equal to 7.0-7.2). The unexpected finding that the brain forms are more basic than the muscle-specific enzyme is supported by agarose and cellulose acetate electrophoresis and ion-exchange chromatography properties.     

Relation of growth process to spatial patterns of electric potential and enzyme activity in bean roots

The electric spatial pattern and invertase activity distribution in growing roots of azuki bean (Phaseolus chrysanthos) have been studied. The electric potential near the surface along the root showed a banding pattern with a spatial period of about 2 cm. It was found that the enzyme activity has a peak around 3-7 mm from the root tip, in good agreement with the position of the first peak of the electric potential, which is located a little behind the elongation zone. An inhomogeneous distribution of ATP content was also detected along the root. Experiments on the electric isolation of the elongation zone from the mature zone and acidification treatment showed that H+ is transported from the mature-side to elongation-side regions, causing tip elongation through an acid-growth mechanism. Both acidification and electric disturbance on growing roots affected growth significantly. Simultaneous measurements of electric potential and enzyme activity clearly showed a good correlation between these two quantities and growth speed. From an analogy with the Characean banding, the spatio-temporal organization via the cell membrane in electric potential and enzyme activity can be regarded as a dissipative structure arising far from equilibrium. These experimental results can be interpreted with a new mechanism that the dissipative structure is formed spontaneously along the whole root, accompanied by energy metabolism, to make H+ flow into the root tip.     

Correlation between dielectric property by dielectrophoretic levitation and growth activity of cells exposed to electric field

The purpose of this study is to develop a system analyzing cell activity by the dielectrophoresis method. Our previous studies revealed a correlation between the growth activity and dielectric property (Re[K(ω)]) of mouse hybridoma 3-2H3 cells using dielectrophoretic levitation. Furthermore, it was clarified that the differentiation activity of many stem cells could be evaluated by the Re[K(ω)] without differentiation induction. In this paper, 3-2H3 cells exposed to an alternating current (AC) electric field or a direct current (DC) electric field were cultivated, and the influence of damage by the electric field on the growth activity of the cells was examined. To evaluate the activity of the cells by measuring the Re[K(ω)], the correlation between the growth activity and the Re[K(ω)] of the cells exposed to the electric field was examined. The relations between the cell viability, growth activity, and Re[K(ω)] in the cells exposed to the AC electric field were obtained. The growth activity of the cells exposed to the AC electric field could be evaluated by the Re[K(ω)]. Furthermore, it was found that the adverse effects of the electric field on the cell viability and the growth activity were smaller in the AC electric field than the DC electric field.