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Uninterrupted microscopic observation and real-time imaging of cell behavior during exposure to the stimulus, for example, electric and/or magnetic fields, especially for periods of several days, has been a challenge in experimental bioelectromagnetics due to a lack of proper gas/temperature conditions outside the incubator. Conventional mini-incubators might suffer from stray fields produced by heating elements. We report an in vitro electric and magnetic fields (EMF) exposure system embedded inside a novel under-the-microscope mini-CO2 -incubator with a unique design to avoid electromagnetic interference from the heating and circulation functions while ensuring the requisite temperature. A unique, reconfigurable array of electrodes and/or coils excited by calculated current distributions among array elements is designed to provide excellent field uniformity and controllable linear or circular polarization (even at very low frequencies) of the EMF within the cell culture. Using standard biochemical assays, long-term cell viability has been verified and compared with a conventional incubator. Cell orientation/migration in three-dimensional culture made of collagen-hydrogels has been successfully observed in vitro, in long-term, and in real-time under the influence of DC electric fields with the device.
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Campos Electromagnéticos , Campos Magnéticos , Incubadoras , Temperatura , ElectricidadRESUMEN
Electrical stimulation of peripheral nerve fibers has always been an attractive field of research. Due to the higher activation threshold, the stimulation of small fibers is accompanied by the stimulation of larger ones. It is therefore necessary to design a specific stimulation theme in order to only activate narrow fibers. There is evidence that stimulating Aδ fibers can activate endogenous pain-relieving mechanisms. However, both selective stimulation and reducing pain by activating small nociceptive fibers are still poorly investigated. In this study, using high-frequency stimulation waveforms (5-20 kHz), computational modeling provides a simple framework for activating narrow nociceptive fibers. Additionally, a model of myelinated nerve fibers is modified by including sodium-potassium pump and investigating its effects on neuronal stimulation. Besides, a modified mathematical model of pain processing circuits in the dorsal horn is presented that consists of supraspinal pain control mechanisms. Hence, by employing this pain-modulating model, the mechanism of the reduction of pain by activating nociceptive fibers is explored. In the case of two fibers with the same distance from the point source electrode, a single stimulation waveform is capable of blocking one large fiber and stimulating another small fiber. Noteworthy, the Na/K pump model demonstrated that it does not have a significant effect on the activation threshold and firing frequency of fiber. Ultimately, results suggest that the descending pathways of Locus coeruleus may effectively contribute to pain relief through stimulation of nociceptive fibers, which will be beneficial for clinical interventions.
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Neuronas , Dolor , Humanos , Estimulación Eléctrica , Simulación por ComputadorRESUMEN
Fracture healing happens naturally in most bone break cases. Occasionally prolongation of restoration period or non-union of the fracture may occur, where electrical stimulation has been shown to facilitate bone restoration by stimulating osteoblasts. Despite clinical use, a comprehensive computational model linking the applied currents to the stimulating field in the fracture has been missing. In this paper, we investigate the input current needed to stimulate osteoblasts in a fracture in the human forearm. Optimal current is computed for various fracture configurations, and sensitivity to frequency and inter/intrapersonal variance in dielectric properties are analyzed. Stimulation thresholds at the fracture site are based on detailed review of experimental studies. Our results show that for a 1 mm thick 30° fracture with a 15 Hz sinusoidal field, the input current amounts to a maximum of 3.77 µA. Minimum and maximum required current levels are plotted versus fracture parameters, all of which comply with the ICNIRP standard. Simulation results are supported by several experimental reports. Our model is useful for understanding the effects of various geometrical and electrical factors on clinical outcome, and serves as a theoretical aid in the design of more efficient systems. Bioelectromagnetics. 40:128-135, 2019. © 2019 Bioelectromagnetics Society.
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Terapia por Estimulación Eléctrica/instrumentación , Curación de Fractura/efectos de la radiación , Fracturas Óseas/terapia , Variación Biológica Poblacional , Huesos , Humanos , Modelos Biológicos , Osteoblastos/efectos de la radiaciónRESUMEN
In this paper, morphological effects of electric fields on avian erythrocytes (nucleated red blood cells) have been studied in detail. Morphological changes include rounding and cytoplasm transparency. It has been shown that the effect is non-thermal. Careful imaging and image analyses have been carried out to show that the degree of this effect is frequency-dependent, and has a higher conversion rate at higher temperatures. Furthermore, to better understand the mechanisms behind the morphological changes, we investigated the dedifferentiation hypothesis and performed a series of tests on avian erythrocytes including fluorescence spectroscopy for hemoglobin, and tests on human umbilical cord blood, mesenchymal stem cells, and bone marrow mesenchymal stem cells including flow-cytometry analysis for expression of certain markers and calcium staining. Bioelectromagnetics. 2019;40:375-390. © 2019 Bioelectromagnetics Society.
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Campos Electromagnéticos , Eritrocitos/citología , Sangre Fetal/citología , Células Madre Mesenquimatosas/citología , Animales , Aves , Diferenciación Celular , Línea Celular , Estimulación Eléctrica , Humanos , Concentración de Iones de Hidrógeno , TemperaturaRESUMEN
In recent years, microscopy has revolutionized the study of dynamic living cells. However, performing long-term live cell imaging requires stable environmental conditions such as temperature, pH, and humidity. While standard incubators have traditionally provided these conditions, other solutions, like stagetop incubators are available. To further enhance the accessibility of stable cell culture environments for live cell imaging, we developed a portable CO2 cell culture mini-incubator that can be easily adapted to any x-y inverted microscope stage, enabling long-term live cell imaging. This mini-incubator provides and maintains stable environmental conditions and supports cell viability comparable to standard incubators. Moreover, it allows for parallel experiments in the same environment, saving both time and resources. To demonstrate its functionality, different cell lines (VERO and MDA-MB-231) were cultured and evaluated using various assays, including crystal violet staining, MTT, and flow cytometry tests to assess cell adhesion, viability, and apoptosis, respectively. Time-lapse imaging was performed over an 85-h period with MDA-MB-231 cells cultured in the mini-incubator. The results indicate that this device is a viable solution for long-term imaging and can be applied in developmental biology, cell biology, and cancer biology research where long-term time-lapse recording is required.
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Dióxido de Carbono , Técnicas de Cultivo de Célula , Imagen de Lapso de Tiempo , Técnicas de Cultivo de Célula/métodos , Incubadoras , Línea CelularRESUMEN
A four-electrode system and high-voltage amplitude stimulation (with a voltage of more than 1 V) are always used for nonlinear spectroscopy of biological solutions like yeast suspension. Multiple studies indicate that yeast membrane activities are the primary source of the harmonics detected from yeast suspensions. However, other investigations have demonstrated that at least part of the harmonics are mainly due to the nonlinear impedance of the electrode-electrolyte interface. Here, we want to find a mathematical model to analyze the odd harmonics generated by the interface. The KCl solution is the basis of the essential medium used in yeast culture. No study models the nonlinear behavior of the KCl solution using four-electrode. We used a sinusoidal voltage with a low frequency and a high amplitude to test an ionic solution with 1, 30, and 130 mM of potassium chloride. The voltage had no direct current component. Here, we show that the relationship between the amplitude of the excitation voltage and the third harmonic of the current can be accurately represented using the Butler-Volmer equation. Additionally, we analyze the impact of potassium chloride (KCl) concentration on the behavior of the third harmonic. This paper presents a method and model that can be used for nonlinear spectroscopy studies of biological solutions using a four-electrode system with the aim of deriving information from cells without considering the interface effect.
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We designed a rectangular waveguide exposure system to study the effects of mobile phone frequency (940 MHz) electromagnetic fields (EMF) on luciferase structure and activity. The luciferase activity of exposed samples was significantly higher than that of unexposed samples. Dynamic light scattering of the exposed samples showed smaller hydrodynamic radii compared to unexposed samples (20 nm vs. 47 nm ± 5%). The exposed samples also showed less tendency to form aggregates, monitored by turbidity measurements at l = 360 nm. A microwave dielectric measurement was performed to study the hydration properties of luciferase solutions with a precision network analyzer over frequency ranges from 0.2 to 20 GHz before and after exposure. The change in the dielectric properties of the exposed luciferase solution was related to the disaggregation potency of the applied field. Together, our results suggested that direct interactions with luciferase molecules and its dipole moment were responsible for the reduced aggregation and enhanced luciferase activity upon exposure to the EMF.
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Campos Electromagnéticos , Luciferasas/metabolismo , Luciferasas/efectos de la radiación , Animales , Espectroscopía Dieléctrica , Cinética , Conformación Proteica/efectos de la radiaciónRESUMEN
Introduction: Transcranial direct current stimulation (tDCS) has been studied as an adjunctive treatment option for substance use disorders (SUDs). Alterations in brain structure following SUD may change tDCS-induced electric field (EF) and subsequent responses; however, group-level differences between healthy controls (HC) and participants with SUDs in terms of EF and its association with cortical architecture have not yet been modeled quantitatively. This study provides a methodology for group-level analysis of computational head models to investigate the influence of cortical morphology metrics on EFs. Methods: Whole-brain surface-based morphology was conducted, and cortical thickness, volume, and surface area were compared between participants with cannabis use disorders (CUD) (n=20) and age-matched HC (n=22). Meanwhile, EFs were simulated for bilateral tDCS over the dorsolateral prefrontal cortex. The effects of structural alterations on EF distribution were investigated based on individualized computational head models. Results: Regarding EF, no significant difference was found within the prefrontal cortex; however, EFs were significantly different in left-postcentral and right-superior temporal gyrus (P<0.05) with higher levels of variance in CUD compared to HC [F(39, 43)=5.31, P<0.0001, C=0.95]. Significant differences were observed in cortical area (caudal anterior cingulate and rostral middle frontal), thickness (lateral orbitofrontal), and volume (paracentral and fusiform) between the two groups. Conclusion: Brain morphology and tDCS-induced EFs may be changed following CUD; however, differences between CUD and HCs in EFs do not always overlap with brain areas that show structural alterations. To sufficiently modulate stimulation targets, whether individuals with CUD need different stimulation doses based on tDCS target location should be checked.
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The coronavirus disease caused by the SARS-CoV-2 virus has affected people worldwide for more than two years. Here we present a new diagnostic method based on nonlinear dielectric spectroscopy to detect the presence of the SARS-CoV-2 virus in swab samples. A known current is injected into the virus sample suspension, and the biomarker is the third harmonic detected in the power spectrum of the recorded signal. Computational modeling of harmonic production supports the hypothesis of ion channels (the E-protein) with nonlinear current-voltage characteristics being present on the virus envelope as a possible origin of harmonics. The developed system is able to distinguish between positive and negative samples with 5-10 dBc (decibels relative to the carrier) higher third harmonic ratios in positive samples, in agreement with the computational estimation. Our early results demonstrate that this method can detect the virus in solution. This is the first time harmonic signatures are used to detect SARS-CoV-2 in swab samples.
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Técnicas Biosensibles , COVID-19 , COVID-19/diagnóstico , Espectroscopía Dieléctrica , Humanos , SARS-CoV-2 , Manejo de EspecímenesRESUMEN
Two challenges to optimizing transcranial direct current stimulation (tDCS) are selecting between, often similar, electrode montages and accounting for inter-individual differences in response. These two factors are related by how tDCS montage determines current flow through the brain considered across or within individuals. MRI-based computational head models (CHMs) predict how brain anatomy determines electric field (EF) patterns for a given tDCS montage. Because conventional tDCS produces diffuse brain current flow, stimulation outcomes may be understood as modulation of global networks. Therefore, we developed a network-led, rather than region-led, approach. We specifically considered two common "frontal" tDCS montages that nominally target the dorsolateral prefrontal cortex; asymmetric "unilateral" (anode/cathode: F4/Fp1) and symmetric "bilateral" (F4/F3) electrode montages. CHMs of 66 participants were constructed. We showed that cathode location significantly affects EFs in the limbic network. Furthermore, using a finer parcellation of large-scale networks, we found significant differences in some of the main nodes within a network, even if there is no difference at the network level. This study generally demonstrates a methodology for considering the components of large-scale networks in CHMs instead of targeting a single region and specifically provides insight into how symmetric vs asymmetric frontal tDCS may differentially modulate networks across a population.
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Encéfalo/fisiología , Red Nerviosa/fisiología , Estimulación Transcraneal de Corriente Directa , Adulto , Humanos , Masculino , Corteza Prefrontal/fisiologíaRESUMEN
A highly sensitive impedimetric bio-chip based on vertically aligned multiwall carbon nanotubes (VAMWCNTs), was applied in direct interaction with lung cancer cells. Our tool provided both inducing and monitoring the bioelectrical changes in the cells initiated by electromagnetic (EM) wave stimulation. EM wave of 940MHz frequency with different intensities was used. Here, wave ablation might accumulate electrical charge on the tips of nanotubes penetrated into cell's membrane. The charge might induce ionic exchanges into the cell and cause alterations in electrical states of the membrane. Transmembrane electrostatic/dynamic states would be strongly affected due to such exchanges. Our novel modality was that, the cells' vitality changes caused by charge inductions were electrically detected with the same nanotubes in the architecture of electrodes for impedance measurement. The responses of the sensor were confirmed by electron and florescent microscopy images as well as biological assays. In summation, our method provided an effective biochip for enhancing and detecting external EM stimulation on the cells useful for future diagnostic and therapeutic applications, such as wave-guided drug-resistance breakage.