Effect of extremely low frequency magnetic fields on oxidative balance in rat brains subjected to an experimental model of chronic unpredictable mild stress.

BACKGROUND: There has been an increasing interest in researching on the effects of extremely low-frequency magnetic fields on living systems. The mechanism of action of extremely low-frequency magnetic fields on organisms has not been established. One of the hypotheses is related to induce changes in oxidative balance. In this study, we measured the effects of chronic unpredictable mild stress induced-oxidative balance of rat's brain exposed to extremely low-frequency magnetic fields. METHODS: A first experiment was conducted to find out if 14 days of chronic unpredictable mild stress caused oxidative unbalance in male Wistar rat's brain. Catalase activity, reduced glutathione concentration, and lipoperoxidation were measured in cerebrum and cerebellum. In the second experiment, we investigate the effects of 7 days extremely low-frequency magnetic fields exposure on animals stressed and unstressed. RESULTS: The main results obtained were a significant increase in the catalase activity and reduced glutathione concentration on the cerebrum of animals where the chronic unpredictable mild stress were suspended at day 14 and then exposed 7 days to extremely low-frequency magnetic fields. Interestingly, the same treatment decreases the lipoperoxidation in the cerebrum. The stressed animals that received concomitant extremely low frequency magnetic fields exposure showed an oxidative status like stressed animals by 21 days. Thus, no changes were observed on the chronic unpredictable mild stress induced-oxidative damage in the rat's cerebrum by the extremely low-frequency magnetic field exposure together with chronic unpredictable mild stress. CONCLUSIONS: The extremely low-frequency electromagnetic field exposure can partially restore the cerebrum antioxidant system of previously stressed animals.

Motor learning impairment in rats under a high sucrose diet.

Motor learning skills are reliable indicators of behavioral acquisition and cognitive disorders. The ease with which learning skills are measured disparities the complexity of the interpretation concerning neural plasticity. Conversely, a wealth of information regarding metabolic derangements has long been reported with direct connection to high sucrose diets. However, the impact of excessive sucrose consumption on undergoing cognitive processes has been only scarcely addressed up to now. Therefore, the goal of this work was to describe the associative relationship between high sucrose consumption and changes in motor learning skills acquisition. Motor learning impairments conditioned by central alterations are hypothesized. Rotarod, elevated plus-maze and open field trials, along with metabolic and pro-inflammatory biomarkers tests in Wistar rats under a high sucrose treatment, were performed. Motor learning impairment in high sucrose diet-treated rats was found while spontaneous locomotor activity remained unchanged. Even though, no anxiety-like behavior under high sucrose diet-treatment was observed. Consistently, the worst outcome in the glucose tolerance test was developed, the worst motor learning performance was observed. Furthermore, insulin resistance correlated positively with a pro-inflammatory state and a decreased latency to fall in the rotarod test. Indeed, C-reactive protein and tumor necrosis factor-α serum levels, along with the homeostasis model assessment of insulin resistance (HOMA-IR), significantly increased in motor learning impairment. Together, these results support behavioral, metabolic and pro-inflammatory changes associated with deleterious changes in central nervous system likely involving crucial motor learning structures. Underlying pro-inflammatory-triggered processes may explain cognitive disorders in advanced states of metabolic derangements.

α5GABAA receptors play a pronociceptive role and avoid the rate-dependent depression of the Hoffmann reflex in diabetic neuropathic pain and reduce primary afferent excitability.

Diabetic neuropathy is an incapacitating complication in diabetic patients. The cellular and molecular mechanisms involved in this pathology are poorly understood. Previous studies have suggested that the loss of spinal GABAergic inhibition participate in painful diabetic neuropathy. However, the role of extrasynaptic α5 subunit-containing GABAA (α5GABAA) receptors in this process is not known. The purpose of this study was to investigate the role of α5GABAA receptors in diabetes-induced tactile allodynia, loss of rate-dependent depression (RDD) of the Hoffmann reflex (HR), and modulation of primary afferent excitability. Intraperitoneal administration of streptozotocin induced tactile allodynia. Intrathecal injection of α5GABAA receptor inverse agonist, L-655,708, produced tactile allodynia in naive rats, whereas it reduced allodynia in diabetic rats. In healthy rats, electrical stimulation of the tibial nerve at 5 Hz induced RDD of the HR, although intrathecal treatment with L-655,708 (15 nmol) abolished RDD of the HR. Streptozotocin induced the loss of RDD of the HR, while intrathecal L-655,708 (15 nmol) restored RDD of the HR. L-655,708 (15 nmol) increased tonic excitability of the primary afferents without affecting the phasic excitability produced by the primary afferent depolarization. α5GABAA receptors were immunolocalized in superficial laminae of the dorsal horn and L4 to L6 dorsal root ganglion. Streptozotocin increased mean fluorescence intensity and percentage of neurons expressing α5GABAA receptors in dorsal horn and L4 to L6 dorsal root ganglia in 10-week diabetic rats. Our results suggest that spinal α5GABAA receptors modulate the HR, play an antinociceptive and pronociceptive role in healthy and diabetic rats, respectively, and are tonically active in primary afferents.

Motor Imagery-Based Brain-Computer Interface Coupled to a Robotic Hand Orthosis Aimed for Neurorehabilitation of Stroke Patients.

Motor imagery-based brain-computer interfaces (BCI) have shown potential for the rehabilitation of stroke patients; however, low performance has restricted their application in clinical environments. Therefore, this work presents the implementation of a BCI system, coupled to a robotic hand orthosis and driven by hand motor imagery of healthy subjects and the paralysed hand of stroke patients. A novel processing stage was designed using a bank of temporal filters, the common spatial pattern algorithm for feature extraction and particle swarm optimisation for feature selection. Offline tests were performed for testing the proposed processing stage, and results were compared with those computed with common spatial patterns. Afterwards, online tests with healthy subjects were performed in which the orthosis was activated by the system. Stroke patients' average performance was 74.1 ± 11%. For 4 out of 6 patients, the proposed method showed a statistically significant higher performance than the common spatial pattern method. Healthy subjects' average offline and online performances were of 76.2 ± 7.6% and 70 ± 6.7, respectively. For 3 out of 8 healthy subjects, the proposed method showed a statistically significant higher performance than the common spatial pattern method. System's performance showed that it has a potential to be used for hand rehabilitation of stroke patients.

Resistive Part of Impedance as a Possible Indicator of Hepatocellular Carcinoma.

BACKGROUND AND AIMS: In this work, the multi-frequency impedance both in normal and liver cancer tissues was studied. This was to investigate the feasibility to detect liver cancer by a low cost, easy to use, and a relatively non-invasive electrical impedance measure technique, and thus potentially improving liver cancer diagnosis. METHODS: Hepatocellular carcinoma (HCC) was induced in male Wistar rats by the administration of diethylnitrosamine (DEN) during 16 weeks. The electrical impedances at a frequency sweep of 10-100 KHz in the whole body and 10-60 KHz in the liver were taken at the end of the treatment. RESULTS: The electrical impedance showed that the real component values of the impedance change in HCC. In addition, we found that the imaginary component was not associated with HCC. CONCLUSION: Our results suggest that the electrical impedance may be used as a diagnostic HCC tool.

Simple and portable low frequency lock-in amplifier designed for photoacoustic measurements and its application to thermal effusivity determination in liquids.

The lock-in amplifier is a very useful instrument for observing very small signals under adverse signal-to-noise conditions. In this work, we describe a simple and portable lock-in amplifier designed to be used in photoacoustic measurements. The device was used to measure the thermal effusivity of eight different liquid samples (distilled water, glycerol, acetone, ethanol, 2-propanol, chloroform, hexane, and methanol), as well as the effusivity of acetone in aqueous solution at distinct concentrations, giving good results. The instrument has a bandwidth of 10 Hz-10 kHz and a sensitivity of 1 µV.

Tonically Active α5GABAA Receptors Reduce Motoneuron Excitability and Decrease the Monosynaptic Reflex.

Motoneurons, the final common path of the Central Nervous System (CNS), are under a complex control of its excitability in order to precisely translate the interneuronal pattern of activity into skeletal muscle contraction and relaxation. To fulfill this relevant function, motoneurons are provided with a vast repertoire of receptors and channels, including the extrasynaptic GABAA receptors which have been poorly investigated. Here, we confirmed that extrasynaptic α5 subunit-containing GABAA receptors localize with choline acetyltransferase (ChAT) positive cells, suggesting that these receptors are expressed in turtle motoneurons as previously reported in rodents. In these cells, α5GABAA receptors are activated by ambient GABA, producing a tonic shunt that reduces motoneurons' membrane resistance and affects their action potential firing properties. In addition, α5GABAA receptors shunted the synaptic excitatory inputs depressing the monosynaptic reflex (MSR) induced by activation of primary afferents. Therefore, our results suggest that α5GABAA receptors may play a relevant physiological role in motor control.

Effect of Intermediate-Frequency Repetitive Transcranial Magnetic Stimulation on Recovery following Traumatic Brain Injury in Rats.

Traumatic brain injury (TBI) represents a significant public health concern and has been associated with high rates of morbidity and mortality. Although several research groups have proposed the use of repetitive transcranial magnetic stimulation (rTMS) to enhance neuroprotection and recovery in patients with TBI, few studies have obtained sufficient evidence regarding its effects in this population. Therefore, we aimed to analyze the effect of intermediate-frequency rTMS (2 Hz) on behavioral and histological recovery following TBI in rats. Male Wistar rats were divided into six groups: three groups without TBI (no manipulation, movement restriction plus sham rTMS, and movement restriction plus rTMS) and three groups subjected to TBI (TBI only, TBI plus movement restriction and sham rTMS, and TBI plus movement restriction and rTMS). The movement restriction groups were included so that rTMS could be applied without anesthesia. Our results indicate that the restriction of movement and sham rTMS per se promotes recovery, as measured using a neurobehavioral scale, although rTMS was associated with faster and superior recovery. We also observed that TBI caused alterations in the CA1 and CA3 subregions of the hippocampus, which are partly restored by movement restriction and rTMS. Our findings indicated that movement restriction prevents damage caused by TBI and that intermediate-frequency rTMS promotes behavioral and histologic recovery after TBI.

Gender Differences in Quantitative Electroencephalogram During a Simple Hand Movement Task in Young Adults.

BACKGROUND: No consensus has been reached regarding the existence of gender differences during motor tasks in electroencephalography. This could lead to misinterpretation of electroencephalography clinical diagnosis and affect the calibration of brain-computer interfaces. OBJECTIVE: To assess whether there are statistically significant gender differences in electroencephalography recorded during hand movements. METHODS: Electroencephalography data were recorded from 18 women and 18 men while performing hand movements and rest. Electroencephalography power was computed for alpha (8-13 Hz), beta (14-30 Hz), and a broader band including alpha and beta (8-30 Hz) using wavelet transform. Statistical analysis was done using a General Linear Model for repeated measurements (α = 0.05). Additionally, topographic maps were computed for each gender. RESULTS: Significant gender differences were found for the rest condition in all analyzed bands. For the hand movement tasks, gender differences were mainly found in the beta band and located in temporoparietal areas. Power decrease observed in topographic maps was located in the centro-parietal areas for females and the centro-frontal areas for males. Additionally, greater power decreases were observed for women in all analyzed frequency bands. CONCLUSION: Electroencephalography parameters used for the diagnosis of neuromotor diseases, as well as for brain-computer interface calibration, must take gender into account.

Voltage-Independent Inhibition of the Tetrodotoxin-Sensitive Sodium Currents by Oxotremorine and Angiotensin II in Rat Sympathetic Neurons.

Tetrodotoxin-sensitive Na(+) currents have been extensively studied because they play a major role in neuronal firing and bursting. In this study, we showed that voltage-dependent Na(+) currents are regulated in a slow manner by oxotremorine (oxo-M) and angiotensin II in rat sympathetic neurons. We found that these currents can be readily inhibited through a signaling pathway mediated by G proteins and phospholipase C (PLC) ß1. This inhibition is slowly established, pertussis toxin-insensitive, partially reversed within tens of seconds after oxo-M washout, and not relieved by a strong depolarization, suggesting a voltage-insensitive mechanism of inhibition. Specificity of the M1 receptor was tested by the MT-7 toxin. Activation and inactivation curves showed no shift in the voltage dependency under the inhibition by oxo-M. This inhibition is blocked by a PLC inhibitor (U73122, 1-(6-{[(17ß)-3-Methoxyestra-1,3,5(10)-trien-17-yl]amino}hexyl)-1H-pyrrole-2,5-dione), and recovery from inhibition is prevented by wortmannin, a PI3/4 kinase inhibitor. Hence, the pathway involves Gq/11 and is mediated by a diffusible second messenger. Oxo-M inhibition is occluded by screening phosphatidylinositol 4,5-bisphosphate (PIP2)-negative charges with poly-l-lysine and prevented by intracellular dialysis with a PIP2 analog. In addition, bisindolylmaleimide I, a specific ATP-competitive protein kinase C (PKC) inhibitor, rules out that this inhibition may be mediated by this protein kinase. Furthermore, oxo-M-induced suppression of Na(+) currents remains unchanged when neurons are treated with calphostin C, a PKC inhibitor that targets the diacylglycerol-binding site of the kinase. These results support a general mechanism of Na(+) current inhibition that is widely present in excitable cells through modulation of ion channels by specific G protein-coupled receptors.

A simple method for fast temperature changes and its application to thermal activation of TRPV1 ion channels.

BACKGROUND: Thermally activated ion channels function as molecular thermometers and participate in other physiological important functions. The mechanism by which they acquire their exquisite temperature sensitivity is unknown and is currently an area of intense research. For this reason, there is a need for diverse methods to deliver controlled temperature stimuli. NEW METHOD: We have developed a simple, inexpensive and reliable method to deliver temperature pulses to small volumes surrounding the recording area, which can be either a patch-clamp pipette containing a cell-free membrane with thermally activated channels or a whole cell attached to a pipette. RESULTS: Here we developed a micro-heater based on resistive heating of a copper filament enclosed in a glass capillary that is capable of delivering fast and localized temperature changes. We validated the performance of the micro-heaters by analyzing the heat-induced activation of TRPV1 thermoTRP channels recorded in inside-out patches and demonstrate the use of the micro-heaters. COMPARISON WITH EXISTING METHOD(S): The micro-heaters we introduce here are compact, easy to fabricate and to operate. In contrast with bulk solution heaters commercially available, our method is extremely affordable and simple to operate. To the best of our knowledge there are no other similar, commercially available heating methods. CONCLUSIONS: The micro-heater method is simple and should provide a straightforward and rapid experimental tool to study mechanisms in thermally activated ion channels.

Extrasynaptic α6 subunit-containing GABAA receptors modulate excitability in turtle spinal motoneurons.

Motoneurons are furnished with a vast repertoire of ionotropic and metabotropic receptors as well as ion channels responsible for maintaining the resting membrane potential and involved in the regulation of the mechanisms underlying its membrane excitability and firing properties. Among them, the GABAA receptors, which respond to GABA binding by allowing the flow of Cl- ions across the membrane, mediate two distinct forms of inhibition in the mature nervous system, phasic and tonic, upon activation of synaptic or extrasynaptic receptors, respectively. In a previous work we showed that furosemide facilitates the monosynaptic reflex without affecting the dorsal root potential. Our data also revealed a tonic inhibition mediated by GABAA receptors activated in motoneurons by ambient GABA. These data suggested that the high affinity GABAA extrasynaptic receptors may have an important role in motor control, though the molecular nature of these receptors was not determined. By combining electrophysiological, immunofluorescence and molecular biology techniques with pharmacological tools here we show that GABAA receptors containing the α6 subunit are expressed in adult turtle spinal motoneurons and can function as extrasynaptic receptors responsible for tonic inhibition. These results expand our understanding of the role of GABAA receptors in motoneuron tonic inhibition.

An approach to improve the performance of subject-independent BCIs-based on motor imagery allocating subjects by gender.

BACKGROUND: One of the difficulties for the implementation of Brain-Computer Interface (BCI) systems for motor impaired patients is the time consumed in the system design process, since patients do not have the adequate physical nor psychological conditions to complete the process. For this reason most of BCIs are designed in a subject-dependent approach using data of healthy subjects. The developing of subject-independent systems is an option to decrease the required training sessions to design a BCI with patient functionality. This paper presents a proof-of-concept study to evaluate subject-independent system based on hand motor imagery taking gender into account. METHODS: Subject-Independent BCIs are proposed using Common Spatial Patterns and log variance features of two groups of healthy subjects; one of the groups was composed by people of male gender and the other one by people of female gender. The performance of the developed gender-specific BCI designs was evaluated with respect to a subject-independent BCI designed without taking gender into account, and afterwards its performance was evaluated with data of two healthy subjects that were not included in the initial sample. As an additional test to probe the potential use for subcortical stroke patients we applied the methodology to two patients with right hand weakness. T-test was employed to determine the significance of the difference between traditional approach and the proposed gender-specific approach. RESULTS: For most of the tested conditions, the gender-specific BCIs have a statistically significant better performance than those that did not take gender into account. It was also observed that with a BCI designed with log-variance features in the alpha and beta band of healthy subjects' data, it was possible to classify hand motor imagery of subcortical stroke patients above the practical level of chance. CONCLUSIONS: A larger subjects' sample test may be necessary to improve the performances of the gender-specific BCIs and to further test this methodology on different patients. The reduction of complexity in the implementation of BCI systems could bring these systems closer to applications such as controlling devices for the motor rehabilitation of stroke patients, and therefore, contribute to a more effective neurological rehabilitation.

[Characterization of electrical brain activity related to hand motor imagery in healthy subjects]. / Caracterización de la actividad eléctrica cerebral relacionada con la imaginacián del movimiento de la mano en sujetos sanos.

Brain computer interface systems (BCI) translate the intentions of patients affected with locked-in syndrome through the EEG signal characteristics, which are converted into commands used to control external devices. One of the strategies used, is to decode the motor imagery of the subject, which can modify the neuronal activity in the sensory-motor areas in a similar way to which it is observed in real movement. The present study shows the activation patterns that are registered in motor and motor imagery tasks of right and left hand movement in a sample of young healthy subjects of Mexican nationality. By means of frequency analysis it was possible to determine the difference conditions of motor imagery and movement. Using U Mann- Whitney tests, differences with statistical significance (p < 0.05) where obtained, in the EEG channels C3, Cz, C4, T3 and P3 in the mu and beta rhythms, for subjects with similar characteristics (age, gender, and education). With these results, it would be possible to define a classifier or decoder by gender that improves the performance rate and diminishes the training time, with the goal of designing a functional BCI system that can be transferred from the laboratory to the clinical application in patients with motor disabilities.

Gß2 mimics activation kinetic slowing of CaV2.2 channels by noradrenaline in rat sympathetic neurons.

Several neurotransmitters and hormones acting through G protein-coupled receptors elicit a voltage-dependent regulation of CaV2.2 channels, having profound effects on cell function and the organism. It has been hypothesized that protein-protein interactions define specificity in signal transduction. Yet it is unknown how the molecular interactions in an intracellular signaling cascade determine the specificity of the voltage-dependent regulation induced by a specific neurotransmitter. It has been suspected that specific effector regions on the Gß subunits of the G proteins are responsible for voltage-dependent regulation. The present study examines whether a neurotransmitter's specificity can be revealed by simple ion-current kinetic analysis likely resulting from interactions between Gß subunits and the channel-molecule. Noradrenaline is a neurotransmitter that induces voltage-dependent regulation. By using biochemical and patch-clamp methods in rat sympathetic neurons we examined calcium current modulation induced by each of the five Gß subunits and found that Gß2 mimics activation kinetic slowing of CaV2.2 channels by noradrenaline. Furthermore, overexpression of the Gß2 isoform reproduces the effect of noradrenaline in the willing-reluctant model. These results advance our understanding on the mechanisms by which signals conveying from a variety of membrane receptors are able to display precise homeostatic responses.

PIP2 hydrolysis is responsible for voltage independent inhibition of CaV2.2 channels in sympathetic neurons.

GPCRs regulate Ca(V)2.2 channels through both voltage dependent and independent inhibition pathways. The aim of the present work was to assess the phosphatidylinositol-4,5-bisphosphate (PIP2) as the molecule underlying the voltage independent inhibition of Ca(V)2.2 channels in SCG neurons. We used a double pulse protocol to study the voltage independent inhibition and changed the PIP(2) concentration by means of blocking the enzyme PLC, filling the cell with a PIP(2) analogue and preventing the PIP(2) resynthesis with wortmannin. We found that voltage independent inhibition requires the activation of PLC and can be hampered by internal dialysis of exogenous PIP(2). In addition, the recovery from voltage independent inhibition is blocked by inhibition of the enzymes involved in the resynthesis of PIP(2). These results support that the hydrolysis of PIP(2) is responsible for the voltage independent inhibition of Ca(V)2.2 channels.

Dissociation of CA3 pyramidal cells with attached, functional, identified mossy fiber and interneuronal boutons for studying glutamatergic and GABAergic synaptic transmission.

Pyramidal cells of CA3 area receive glutamatergic signals from the mossy fibers (MFs), perforant path and collaterals of other pyramidal cells, as well as GABAergic inputs from interneurons. In hippocampal slices, an extracellular stimulation electrode is often used to activate the MFs, with the disadvantage of possibly activating fibers other than MFs. We set-up a preparation that allows the analysis of the glutamatergic input from identified, giant MF boutons as well as of GABAergic inputs from boutons of interneurons on single CA3 pyramidal cells. Mossy fiber boutons were labeled by exposing hippocampal slices to a zinc-reactive fluorescent dye, or by injecting a fluorescent dye in the granule cell layer and allowing its transport along the MFs to their terminals in CA3 area. After conducting an enzyme-free, mechanical dissociation of CA3 area, we obtained pyramidal cells containing fluorescent, giant MF boutons attached to their apical dendrites, as well as boutons of interneuronal origin. Whole cell recordings were then performed, whereby synaptic responses could be evoked by selective stimulation of the identified boutons. The synaptic currents evoked by stimulation of MF boutons, unlike those evoked by stimulation of interneuronal boutons, underwent strong frequency potentiation and were depressed by activation of metabotropic glutamate receptors, which are characteristics of transmission of MF origin. Combination of fluorophores can be used to label different tracts/boutons allowing the study of the different characteristics of neurotransmitter release from a variety of sources on single target cells.

Uncoupling charge movement from channel opening in voltage-gated potassium channels by ruthenium complexes.

The Kv2.1 channel generates a delayed-rectifier current in neurons and is responsible for modulation of neuronal spike frequency and membrane repolarization in pancreatic ß-cells and cardiomyocytes. As with other tetrameric voltage-activated K(+)-channels, it has been proposed that each of the four Kv2.1 voltage-sensing domains activates independently upon depolarization, leading to a final concerted transition that causes channel opening. The mechanism by which voltage-sensor activation is coupled to the gating of the pore is still not understood. Here we show that the carbon-monoxide releasing molecule 2 (CORM-2) is an allosteric inhibitor of the Kv2.1 channel and that its inhibitory properties derive from the CORM-2 ability to largely reduce the voltage dependence of the opening transition, uncoupling voltage-sensor activation from the concerted opening transition. We additionally demonstrate that CORM-2 modulates Shaker K(+)-channels in a similar manner. Our data suggest that the mechanism of inhibition by CORM-2 may be common to voltage-activated channels and that this compound should be a useful tool for understanding the mechanisms of electromechanical coupling.

Effects of acute electromagnetic field exposure and movement restraint on antioxidant system in liver, heart, kidney and plasma of Wistar rats: a preliminary report.

PURPOSE: The aim of the present study was to evaluate the early effects of acute (2 h) exposure to extremely low frequency electromagnetic fields (ELF-EMF), as well as movement restraint (MR) and the combination of both on the antioxidant systems in the plasma, liver, kidney, and heart of rats. MATERIALS AND METHODS: Twenty-four adult male Wistar rats were divided in two groups, restrained and unrestrained. The restrained animals were confined into an acrylic tube for 120 min. Half of the animals of each group were exposed to ELF-EMF (60 Hz, 2.4 mT) during the period of restriction. Immediately after treatment, reduced glutathione (GSH), catalase (CAT), superoxide dismutase (SOD), and thiobarbituric acid reactive substances (TBARS) were measured in tissues. RESULTS: GSH concentration was significantly lower in the heart of all experimental animals when compared to the control group; furthermore, the decrease was higher in the liver of restrained animals. SOD activity was lower in the plasma of restrained and EMF exposed animals compared to unrestrained rats. There were no significant differences in CAT activity and TBARS levels among all the experimental groups vs. the control group. CONCLUSION: Two hours of 60 Hz EMF exposure might immediately alter the metabolism of free radicals, decreasing SOD activity in plasma and GSH content in heart and kidney, but does not induce immediate lipid peroxidation. Oxidative stress induced by movement restraint was stronger than that produced by EMF.

Neurite outgrowth on chromaffin cells applying extremely low frequency magnetic fields by permanent magnets.

BACKGROUND AND AIMS: There is an increasing interest about the effects of electromagnetic fields on health and clinical applications. Electromagnetic fields have been shown to promote differentiation and regeneration of many tissues. The purpose of the present study was to evaluate if a magnetic field (MF) varying in time is able to induce neurite outgrowth in cultured chromaffin cells. For this reason, a stimulation system was developed in order to generate a magnetic field, using permanent magnets as a supply. METHODS: In this investigation we used a pair of permanent ferrite magnets. These were mounted in a mechanical system in which both magnets rotate around a culture Petri dish. The stimulation device was designed at Centro de Investigación y de Estudios Avanzados, Avanzados del IPN, Mexico City. Primary cultures of chromaffin cells were stimulated with a magnetic field of 6.4 mT and 4, 7, 10 or 12Hz (2h daily, during a 7-day period). After treatment, percentage of neurite outgrowth was calculated. RESULTS: Our results show that the magnetic fields produced by rotating permanent magnets induced neurite outgrowth on chromaffin cells at 7 and 10Hz. CONCLUSIONS: The present study provides evidence that MFs varying in time (7 and 10Hz) induce neurite outgrowth in chromaffin cells. These studies will contribute to elucidate the effect of noninvasive MF stimulus in order to apply it in future regeneration therapies. Also, the device designed could be used for different kind of cells and may work as a model for future clinical devices.