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An in vitro comparison of the electrical conducting properties of multiport versus single-port epidural catheters for the epidural stimulation test.

Effective conduction of electricity through a catheter is essential for the success of the epidural stimulation test. In this in vitro study we examined the electrical conductivity of single and multiport epidural catheters (with and without embedded metal elements) after being primed with normal saline. Seven different types of 19-gauge catheters (n = 5), either single-port or multiport catheters, with or without embedded metal elements, were studied. The proximal end of each epidural catheter was connected to the cathode of a nerve stimulator via an electrode adapter. The catheter, primed with normal saline, was placed at the bottom of a syringe filled with 5 different volumes of saline (1, 2, 3, 4 and 5 mL) and attached to an electrode adapter. The voltage of the peripheral nerve stimulator was measured using an oscilloscope. The electrical resistance between the proximal and distal end of the catheter was calculated using Ohm's Law. In catheters without metal elements the electrical resistances were too high to be measured. In catheters that had metal elements, the mean electrical resistances of the same catheter design (single-port or multiport) were similar. However, the electrical resistances of the multiport metal reinforced epidural catheters were significantly lower (P < 0.05) than the single-port metal coil reinforced epidural catheters. The volume of saline in the syringe had no impact on the measured electrical resistances. This study suggests that multiport metal reinforced epidural catheters have low electrical resistances and, thus, are a reasonable alternative to single-port catheters for transmitting sufficient current for performing the epidural stimulation test. On the other hand, epidural catheters without metal elements (single-port or multiport) are not suitable for performing the stimulation test.

Anesthesia, Epidural↗

Orientation and directed migration of cultured corneal epithelial cells in small electric fields are serum dependent.

Reorientation and migration of cultured bovine corneal epithelial cells (CECs) in an electric field were studied. Electric field application was designed to model the laterally directed, steady direct current electric fields which arise in an injured corneal epithelium. Single cells cultured in media containing 10% foetal bovine serum showed significant galvanotropism, reorienting to lie perpendicular to electric field vector with a threshold field strength of less than 100 mV/mm. Cells cultured in serum-free medium showed no reorientation until 250 mV/mm. Addition of EGF, bFGF or TGF-beta 1 singly or in combination to serum free medium significantly restored the reorientation response at low field strengths. Both the mean translocation rate and directedness of cell migration were serum dependent. Cultured in medium with serum or serum plus added EGF, single cells showed obvious cathodal migration at 100 mV/mm. Increasing electric field strength enhanced the cathodal directedness of single cell migration. Supplementing serum free medium with growth factors restored the cathodal directed migration of single cells and highest directedness was found for the combination of EGF and TGF-beta 1. Corneal epithelial sheets also migrated towards the cathode in electric fields. Serum or individual growth factors stimulated CEC motility (randomly directed). Applied fields did not further augment migration rates but added a vector to stimulated migration. Electric fields which are present in wounded cornea interact with other environmental factors and may impinge on CECs migration during wound healing. Therapies which combine the application of growth factors and electric fields may be useful clinically.

Animals↗

Registering the response of tissues exposed to an interferential electric current stimulation.

Electrotherapy is a treatment method in which an electric stimulus is used to achieve physiological changes in the body. When increasing the level of the electric stimulus, three sensory, three motor and a pain response can be found. In this paper the three sensory threshold levels (1st just noticeable weak tingling sensation, 2nd a very fine but definite tingling and 3rd moderate strong tingling) were studied with an interferential electric current stimulus. The interferential electric current can be achieved by summing two alternating currents with little different frequencies with each other. As a current source, the study used a value intended for interferential electric current therapy. The threshold value measurements were performed on 5 days to 6 volunteers. The method of limits is a frequently used technique for determining sensory thresholds where ascending or descending stimuli is used and the observer reports of the appearance or disappearance of the sensation, respectively. A Bekesy's algorithm is a threshold tracking method with a variable stimulus. They were used for controlling the interferential electric current stimulus, and hence, for studying the,thresholds. By using the method of limits, the measurements were done at three sensory threshold levels at interferential frequencies deltaf of 10 Hz, 30 Hz, 50 Hz, and 100 Hz caused by two sine waves, one with a frequency between 2490 Hz and 2400 Hz, and the other constant 2500 Hz. In the method based on Bekésy's algorithm the interferential frequency deltaf was increased from 1 Hz to 100 Hz by changing the first frequency from 2499 Hz to 2400 Hz and the measurement was repeated for all three sensory thresholds. The sensory thresholds for interferential current stimuli seem to be quite unaffected by the interferential electric current frequency among the healthy test persons, and no distinct interferential electric current frequencies at which the threshold values should be measured were found in this study.

Adult↗

Neuromuscular electrical stimulation after anterior cruciate ligament surgery.

This randomized, double blind, controlled trial was designed to determine the effectiveness of neuromuscular electrical stimulation of the knee extensor and flexor muscles in the prevention of muscular weakening after anterior cruciate ligament surgery. Neuromuscular electrical stimulation treatment was given in addition to an early exercise therapy regimen and compared with an early exercise therapy regimen alone. Forty-nine patients after anterior cruciate ligament surgery were assigned randomly either to a neuromuscular electrical stimulation and exercise group, a transcutaneous electrical nerve stimulation as analgesic and exercise group, or an exercise alone group as control. All groups received a standard regimen of rehabilitation after anterior cruciate ligament surgery. The neuromuscular electrical stimulation and transcutaneous electrical nerve stimulation group additionally received electrical stimulation during the first 6 weeks after surgery. Patients were measured for isometric and isokinetic torque in the knee extensor and flexor muscles after 6, 12, and 52 weeks. No statistical difference among groups was observed. It is concluded that neuromuscular electrical stimulation in combination with an early exercise therapy regimen is not significantly more effective in reducing weakening than an early exercise therapy regimen alone after anterior cruciate ligament surgery.

Adult↗

Reconstitution of biological molecular generators of electric current. H+-ATPase.

1. Generation of a transmembrane electric potential difference by oligomycin-sensitive ATPase complex, incorporated into spherical or planar phospholipid membrane, has been demonstrated. To this end, penetrating anion probe and direct voltmeter measurement of electric potential across phospholipid membrane were used. It was found that ATP-induced electric response is sensitive to oligomycin and protonophorous uncouplers. 2. The effect of variations in the phospholipid component of proteoliposomes on the electric generation was studied. It was revealed that the usage of mitochondrial phospholipids and phosphatidylethanolamine allows the highest values of membrane potential to be obtained in the case of ATPase proteoliposomes. In the case of cytochrome oxidase and bacteriorhodopsin proteoliposomes, phosphatidylserine was also shown to be quite suitable. Phosphatidylcholine was absolutely ineffective in all cases. 3. In proteoliposomes, containing both ATPase and bacteriorhodopsin, ATP and light induced generation of the electric field of the same direction. 4. In ATPase + cytochrome oxidase proteoliposomes, ATP hydrolysis and ascorbate oxidation was found to support electric generation of the same direction if cytochrome c was inside vesicles. Oxidation via external cytochrome c resulted in formation of electric field of the direction, opposite to that induced by ATP hydrolysis. 5. The data obtained in experiments with proteoliposomes of different types are discussed. The conclusion is made that conversion of energy of different resources into electric form is a common feature of membraneous energy transducers, which is in agreement with the Mitchellian principle of cellular energetics.

Adenosine Triphosphatases↗

Electrical conduction through nerve and DNA.

The aim of the present study was to analyse electric resistivity at different ambient temperatures between 300 to 20K in the frog sciatic nerve and salmon sperm DNA. When the electrical contacts were leaned just into the sciatic nerve, an increase of the sciatic nerve resistivity was observed for 240 K < T < 300 K and a rise of electrical conductivity was apparent below 240 K. This dependence is generally associated with a semiconductor behaviour. Once the sciatic nerve temperature was driven below 250K, the resistivity abruptly decreased and then at temperatures lower than 234 K, it remained constant and close to one tenth of its ambient temperature value. By contrast, when the electrical contacts were leaned into Salmon sperm DNA, the resistivity remained constant between 300K to 20K, showing a high electrical stability at low temperature. Thus, we report the existence of a new form of electric conductivity in the sciatic nerve at low ambient temperature, which in turn has many electric similarities with inorganic or organic superconductors, whereas temperature failed to alter DNA electrical properties until 20K.

Animals↗

Reconstitution of biological molecular generators of electric current. Bacteriochlorophyll and plant chlorophyll complexes.

1. Electric generation by bacteriochlorophyll reaction center complexes from Rhodospirillum rubrum and by photosystem I complexes from pea chloroplasts has been studied. 2. The methods for the proteoliposome reconstitution from azolectin and bacteriochlorophyll- or plant chlorophyll-containing protein complexes have been elaborated. Light-dependent electric responses of the proteoliposomes were detected using (a) phenyldicarbaundecarborane anion (PCB-) probe and (b) direct measurement by a voltmeter in the proteoliposome-planar phospholipid membrane system. 3. Both PCB- and direct measurements demonstrated that bacteriochlorophyll proteoliposomes are competent in light-dependent electric generation (plus outside proteoliposomes). The photoelectric effect was shown to increase on addition of tetramethyl-p-phenylenediamine (TMPD), CoQ6, and vitamin K3, and to decrease on addition of ferricyanide, o-phenanthroline and a protonophorous uncoupler. Estimation of the photoelectromotive force of the bacteriochlorophyll proteoliposome-planar membrane system gave a value of about 0.2 V. The action spectrum of the photoelectric effect was similar to the absorption spectrum of the bacteriochlorophyll complex. 4. Reconstitution of proteoliposomes containing bacteriochlorophyll centers and bacteriorhodopsin resulted in the system generating an electric field whose direction can be changed by varying the spectral composition of the light: the red light, exciting bacteriochlorophyll, induces negative, and the green light, exciting bacteriorhodopsin, induces positive charging of the proteoliposome interior. 5. Association of isolated R. rubrum chromatophores with planar phospholipid membrane was found to give a system demonstrating light-induced electric generation as high as 215 mV in the presence of napthoquinone, TMPD (or phenazine methosulfate, PMS), and ascorbate. Under the same conditions, addition of inorganic pyrophosphate or ATP results in formation of an electric field of the same direction as that induced by light. 6. Proteoliposomes with plant chlorophyll complexes of photosystem I demonstrated light-induced PCB- responses indicating formation of the electric field with plus inside vesicles. The effect required PMS addition. A protonophorous uncoupler and o-phenanthroline were inhibitory. Electric responses in the chlorophyll proteoliposome-planar membrane system were very small (not higher than 10 mV).

Bacterial Chromatophores↗

Middle-latency responses. I. Electrical and acoustic excitation.

The electrically evoked auditory brain-stem response has been used in the past to assess auditory system function with regard to cochlear prosthesis application. The brief latency of the response makes it susceptible to electrical artifact contamination, and waveform identification is often difficult. As a possible alternative for a noninvasive measure of system excitability, the middle-latency response (MLR), elicited by electrical stimulation, was investigated. Middle-latency responses were recorded in response to acoustic and round-window electrical stimulation in albino guinea pigs. Acoustic and electrically evoked MLR waveforms were similar, as were their respective latency/intensity functions. Amplitude/intensity functions for the electric MLR showed greater variability than acoustically evoked MLR functions. The electric MLR is readily evoked and relatively insensitive to electrical artifact in the guinea pig. It is potentially a useful tool in assessing the integrity of auditory pathways and consequently in the development of diagnostic tests for cochlear implant candidates.

Acoustic Stimulation↗

Neurotrophins enhance electric field-directed growth cone guidance and directed nerve branching.

Neurotrophins play major roles in the developing nervous system in controlling neuronal differentiation, neurite outgrowth, guidance and branching, synapse formation and maturation, and neuronal survival or death. There is increasing evidence that nervous system construction takes place in the presence of dc electric fields, which fluctuate dynamically in space and time during embryonic development. These have their origins in the neural tube itself, as well as in surrounding skin and gut. Early disruption of these endogenous electric fields causes failure of the nervous system to form, or else it forms aberrantly. Nerve growth, guidance, and branching are controlled tightly during pathway construction and in vitro dc electric fields have profound effects on each of these behaviours. We have used cultured neurones to ask whether neurotrophins and dc electric fields might interact in shaping neuronal growth, given their coexistence in vivo. Electric field-directed nerve growth generally was enhanced by the simultaneous presentation of several neurotrophins to the growth cone. Under certain circumstances, more nerves turned cathodally, they turned faster, further, and in lower field strengths. Intriguingly, other combinations of dc electric field and neurotrophins (low field strength and neurotrophin 3 (NT-3) switched the direction of growth cone turning. Additionally, cathodally directed nerve growth was faster and directed branching was much more common when electric fields and neurotrophins interacted with neuronal growth cones. Given such profound changes in growth cone behaviour in vitro, neurotrophins and endogenous electric fields are likely to interact in vivo.

Animals↗

Electric field exposure alters serum melatonin but not pineal melatonin synthesis in male rats.

Sprague-Dawley male rats, maintained in a 14:10 h light:dark cycle were exposed for 30 days (starting at 56 days of age) to a 65 kV/m, 60 Hz electric field or to a sham field for 20 h/day beginning at dark onset. Pineal N-acetyltransferase (NAT), hydroxy-indole-o-methyl transferase (HIOMT), and melatonin as well as serum melatonin were assayed. Preliminary data on unexposed animals indicated that samples obtained 4 h into the dark period would reveal either a phase delay or depression in circadian melatonin synthesis and secretion. Exposure to electric fields for 30 days did not alter the expected nighttime increase in pineal NAT, HIOMT, or melatonin. Serum melatonin levels were also increased at night, but the electric field-exposed animals had lower levels than the sham-exposed animals. Concurrent exposure to red light and the electric field or exposure to the electric field at a different time of the day-night period did not reduce melatonin synthesis. These data do not support the hypothesis that chronic electric field exposure reduces pineal melatonin synthesis in young adult male rats. However, serum melatonin levels were reduced by electric field exposure, suggesting the possibility that degradation or tissue uptake of melatonin is stimulated by exposure to electric fields.

Acetylserotonin O-Methyltransferase↗

Effect of electrical charges and fields on injury and viability of airborne bacteria.

In this study, the effects of the electric charges and fields on the viability of airborne microorganisms were investigated. The electric charges of different magnitude and polarity were imparted on airborne microbial cells by a means of induction charging. The airborne microorganisms carrying different electric charge levels were then extracted by an electric mobility analyzer and collected using a microbial sampler. It was found that the viability of Pseudomonas fluorescens bacteria, used as a model for sensitive bacteria, carrying a net charge from 4100 negative to 30 positive elementary charges ranged between 40% and 60%; the viability of the cells carrying >2700 positive charges was below 1.5%. In contrast, the viability of the stress-resistant spores of Bacillus subtilis var. niger (used as simulant of anthrax-causing Bacillus anthracis spores when testing bioaerosol sensors in various studies), was not affected by the amount of electric charges on the spores. Because bacterial cells depend on their membrane potential for basic metabolic activities, drastic changes occurring in the membrane potential during aerosolization and the local electric fields induced by the imposed charges appeared to affect the sensitive cells' viability. These findings facilitate applications of electric charging for environmental control purposes involving sterilization of bacterial cells by imposing high electric charges on them. The findings from this study can also be used in the development of new bioaerosol sampling methods based on electrostatic principles.

Bacillus subtilis↗

Thermomechanical facilitation of swallowing evoked by electrical nerve stimulation in cats.

Application of a cold metal probe to the anterior faucial pillar has been reported to improve swallowing in some patients with dysphagia. Although a variety of stimuli contribute to the initiation of swallowing, the effects of a controlled, cold-thermal stimulus combined with mechanical stimulation have not been examined. It is known that simultaneous stimulation of the glossopharyngeal nerve (IX) and the superior laryngeal nerve may summate to facilitate swallowing in the cat. The goal of this study was to determine whether thermomechanical stimulation of the mucosa innervated by IX would interact with threshold electrical stimulation of the internal laryngeal nerve (ILN) to augment the swallowing response in cats. Four experimental conditions were tested over 24 trials in 4 pentobarbital-anesthetized cats. These included electrical stimulation of ILN, mechanical stimulation of the anterior faucial pillar with a thermode at ambient (room) temperature, concurrent ambient-mechanical and electrical stimulation, and concurrent cold-mechanical and electrical stimulation. Tissue was cooled to 8.9 degrees C during cold-mechanical-electrical stimulation and 25.3 degrees C during ambient-mechanical-electrical and ambient-mechanical alone stimulation. Ambient-mechanical stimulation alone did not produce swallowing. However, both forms of thermomechanical-electrical stimulation elicited a significantly greater number of swallows than did electrical stimulation alone. Therefore, mechanical stimulation with a thermode was capable of modifying the swallowing response in neurologically intact cats.(ABSTRACT TRUNCATED AT 250 WORDS)

Anesthesia↗

Ionic currents that contribute to a sexually dimorphic communication signal in weakly electric fish.

Weakly electric fish produce a communication signal, the electric organ discharge, that is species specific, and in many species, sexually dimorphic. Because the neural circuit that controls the electric organ discharge is relatively simple, it is an excellent model in which to study both the biophysical mechanisms underlying a rhythmic behavior and the neuroendocrine control of a sexually dimorphic behavior. By studying the effects of ion channel blockers on neurons in the medullary pacemaker nucleus, I pharmacologically characterized three ionic currents that influence the pacemaker rhythm, and thus electric organ discharge frequency, in the gymnotiform fish, Apteronotus leptorhynchus. These currents included a tetrodotoxin-sensitive sodium current; a potassium current that was sensitive to 4-aminopyridine; and a calcium current that was sensitive to nickel and cadmium, but resistant to specific blockers of L-, N-, P-, and Q-type calcium currents. The pharmacological profiles of the ionic currents in the pacemaker nucleus are similar to those of ionic currents involved in pacemaking in other neuronal oscillators. Because these ionic currents dramatically influence pacemaker firing frequency, which is directly related to electric organ discharge frequency, these ionic currents are likely targets of steroid hormone action in producing sexual dimorphisms in electric organ discharge frequency. Additional studies are needed to determine how these ionic currents interact to generate the electric organ discharge rhythm and to investigate the possibility that sexual dimorphism in the electric organ discharge results from the actions of gonadal steroids on these ionic currents.

Animal Communication↗

Electrophoretic repatterning of charged cytoplasmic molecules within tissues coupled by gap junctions by externally applied electric fields.

Ionic currents and cytoplasmic voltage gradients have been observed in a variety of polarizing cells and developing tissues. In certain cases, it has been determined that these endogenous electric fields can segregate intracellular charged molecules by electrophoresis; in other cases, the endogenous fields are suspected to have such an influence. Separate theoretical suggestions have been made that extracellular electric currents, whether from a biological or a nonbiological source, should be able to electrophorese intracellular molecules after being conducted through cell membranes into the interior of long single cells [L.F. Jaffe and R. Nuccitelli (1977) Annu. Rev. Biophys. Bioeng. 6, 445-476] or extended ensembles of cells coupled electrotonically by gap junctions [M.S. Cooper (1984) J. Theor. Biol. 111, 123-130]. To test whether external electric fields could redistribute intracellular molecules within a tissue coupled by gap junctions, and to quantitatively measure in situ the electrophoretic mobility of a charged intracellular molecule, we injected 6-carboxyfluorescein into the electrotonically coupled lateral giant neurons of the crayfish abdominal nerve cord. When a dc electric field (0.2-3.4 V/cm) was subsequently applied along the length of the cord, the negatively charged fluorescent dye was observed to migrate through both the cytoplasms and the gap junctions of the lateral giant neurons, toward the anode, at a rate directly proportional to the applied electric field strength (electrophoretic mobility = -0.92 +/- 0.27 micron/sec per V/cm). These results suggest that electric fields of a sufficient magnitude, whether of an exogenous or an endogenous origin, can repattern the distribution of charged molecules within the cytoplasm of an extended ensemble of coupled cells. In addition, these results suggest that externally applied electric fields might be used in studies of pattern formation to repattern the intercellular distribution of charged molecules that are permeant to gap junctions within electrically coupled tissues.

Animals↗

Schistosoma mansoni: effects of bromolysergic acid diethylamide, verapamil, and Ca2+-free solution on the motor activity of the isolated male worm induced by electrical stimulation and oxamniquine.

Electrical stimulation and oxamniquine effect the motor activity of isolated Schistosoma mansoni. Electrical stimulation produced contractions that increased with stimulus intensity. Oxamniquine (10(-4) M) produced an increase in basal tonus and in the frequency and amplitude of the worm's spontaneous contractions. Incubation in the absence of calcium produced a decrease in the basal tonus, abolished the spontaneous contractions of S. mansoni, and abolished the mechanical response induced by electrical stimulation and oxamniquine. The effects of electrical stimulation and oxamniquine were, respectively, significantly reduced and abolished by 10(-6) M verapamil (a calcium channel blocking agent). Bromolysergic acid diethylamide (3 X 10(-5) M), a serotonin blocking agent, reduced the motor response induced by high intensity electrical stimulation and blocked the response induced by oxamniquine. The effects of low intensity electrical stimulation were not modified in the presence of bromolysergic acid. We think that external Ca2+ is important for basal tonus, for spontaneous motor activity, and for motor responses of S. mansoni induced by electrical stimulation and oxamniquine. Serotonin may be important for mechanical responses induced by high intensity electrical stimulation and for responses induced by oxamniquine.

Animals↗

Hair cell mediated responses of the auditory nerve to sinusoidal electrical stimulation of the cochlea in the cat.

Electrical stimulation of the cochlea elicits discharges of auditory nerve fibres which are mediated by the electrical or mechanical stimulation of inner hair cells (electrophonic responses). In order to isolate hair-cell mediated responses from those elicited by electrical stimulation of the nerve, the compound action potential (CAP) evoked by an acoustic probe was forward-masked by sinusoidal monopolar, or localized bipolar electrical stimulation of the base of the cochlea. The degree of masking of a given probe estimated the synaptically mediated response to the masker of the population of auditory nerve fibres innervating the cochlear location tuned to the probe. There was a peak of masking for probes close to the frequency of the electrical stimulus, suggesting a spatial tuning of the hair cell mediated response along the cochlea. This is consistent with excitation of the inner hair cells by a propagating mechanical response which is generated within the electrical field at the base of the cochlea. Furthermore, tuning curves for masking of a given probe were sharply tuned to electrical stimulation close to the probe frequency. This masking was not dependent upon the presence of functional outer hair cells close to the electrodes, suggesting an alternate transduction of electrical to mechanical energy.

Action Potentials↗

Early epidermal changes in heat- and electrically injured pig skin. I. A light microscopic study.

In order to find methods applicable for disclosing electrical torture, pig skin was exposed to heat and electricity under controlled circumstances. Biopsies were obtained immediately after exposure and the morphology of the heat lesions was compared to that of electrical lesions. The cytoplasm of epidermal cells in heat lesions appeared granular or fibrillar, while the nuclei were rather unaffected. In electrical lesions the cytoplasm appeared homogeneous often with a peculiar white colour in hematoxylin-eosin stained sections. The nuclei were either dark and shadowy or enlarged and vesicular. The keratin in electrical lesions often showed a bright yellow colour. Small defects in the epidermis were seen in some electrical lesions. Thus the morphology of electrical lesions differed markedly from that of heat lesions. Although some of these differences may be due to differences in distribution and intensity of energy, it is probable that pH shifts in the cells due to electrolysis is the main cause of the specific morphology of electrical lesions.

Animals↗

A series of models of non-thermal high voltage electrical injuries.

Although many models of electrical injury have been established, none of them are completely typical of the clinical features of electrical injury. As a result, research based on these models were incapable of explaining many clinical phenomena such as continuous tissue necrosis and also were unable to cope with high ratio of amputation of extremities. In order to investigate the mechanism of electrical injuries and better model the condition with similar clinical characteristics we developed a new model. Seventy-five New Zealand rabbits were employed in this study, of them 45 were used in a preliminary experiment including the selection of the size of electrode plate area, damaged extent, time length of electrical injury and interval length between two injuries and so on. Another 30 rabbits were equally divided into five groups with electrical injury times of 6, 12, 18, 24 and 30 cycles, respectively. Observations were made using clinical anatomical exploration, with quantification using an IDBI scale on the 2nd, 8th, 24th, 48th hours and 5th, 15th days, and TC-99m-DMP isotope scanning and gamma photography at 2nd hour and 5th day in post-injury, respectively. The results showed the effective electric field strength was 17,000 V/m, mean current intensity was 554 mA, average current density was 137 mA/cm(2) beneath the small electrode plate with 21 mA/cm(2) beneath big one, and average increase of tissue temperature was 1.73 degrees C during injury process which excluded the possibility of thermal injury. One single wound injury beneath the small plate of the experimental rabbits with loss of injured extremities from 5th to 15th post-injury days in groups 3-5 and obviously progressive tissue necrosis in and outside the wounds were obtained. A series of electrical injured models from mild, moderate, severe, extra severe, and destructive which was exactly similar to the clinical features of electrical injury cases was established.

Animals↗