Electric field driven jetting: an emerging approach for processing living cells

This paper reports for the first time the ability to process living cellular materials by means of electrified jets at electric field strengths of up to 2 kV/mm. Bio-suspensions containing living human Jurkat cells at different concentrations were processed via this jetting approach. The jetting process was carried out at an electric field strength between 0.67 kV/mm and 2 kV/mm, corresponding to an applied voltage of 10-30 kV between two electrodes approximately 15 mm apart. The Jurkat cells were jetted under sterile conditions, collected in petri dishes and incubated for 24 and 48 hours. During and after incubation, cells were assessed for survival and structural damage; cells were found to be unharmed and to retain their integrity under all electric field strengths examined. At all field strengths jetting took place in the unstable mode. Good correlation was observed between droplet distribution plots generated by way of laser spectroscopy and estimated values from measurements of droplet relics.     

Relationship between field strength and arousal response in mice exposed to 60-Hz electric fields

White-footed mice, Peromyscus leucopus, were exposed to 60-Hz electric fields to study the relationship between field strength and three measures of the transient arousal response previously reported to occur with exposures at 100 kV/m. Five groups of 12 mice each were given a series of four 1-h exposures, separated by an hour, with each group exposed at one of the following field strengths: 75, 50, 35, 25, and 10 kV/m; 8 additional mice were sham-exposed with no voltage applied to the field generator. All mice were experimentally naive before the start of the experiment, and all exposures occurred during the inactive (lights-on) phase of the circadian cycle. The first exposure produced immediate increases in arousal measures, but subsequent exposures had no significant effect on any measure. These arousal responses were defined by significant increases of gross motor activity, carbon dioxide production, and oxygen consumption, and were frequently recorded with field strengths of 50 kV/m or higher. Significant arousal responses rarely occurred with exposures at lower field strengths. Responses of mice exposed at 75 and 50 kV/m were similar to previously described transient arousal responses in mice exposed to 100-kV/m electric fields. Less than half of the mice in each of the field strength groups below 50 kV/m showed arousal responses based on Z (standard) scores, but the arousals of the mice that did respond were similar to those of mice exposed at higher field strengths. Polynomial regression was used to calculate the field strength producing the greatest increases for each of the arousal measures. The results show that the amplitude of the transient arousal response is related to the strength of the electric field, but different measures of arousal may have different relationships to field strength.     

Electric properties and structure of DNA-restriction fragments from measurements of the electric dichroism

The electric dichroism of 17 homogeneous DNA fragments, ranging in size from 43 to 4362 base-pairs, has been analyzed in high electric fields. The orientation of the small fragments can be described in terms of an induced dipole moment, whereas the large fragments are oriented according to a constant dipole mechanism. In the intermediate size range, DNA orients according to an induced dipole mechanism at low field strengths and according to a constant dipole mechanism at high field strengths. From these observations we propose an orientation mechanism with a saturating induced dipole. The induced dipole observed at low field strengths is saturated at a field strength Eo within a transition range Em to give a constant dipole moment at high field strengths. These parameters together with the polarizability and the limit reduced dichroism are evaluated by a least-squares analysis of the experimental data. Eo and Em are found to decrease with increasing chain length from Eo approximately 40 kV/cm (Em approximately 14 kV/cm) at 65 base-pairs to 10 kV/cm (6 kV/cm) at 194 base-pairs. The polarizability is found to increase with the square of the chain length, whereas the saturated dipole increases with chain length N at low N and goes to a limit value at high N. The temperature dependence of the orientation parameters is found to be very small. The values obtained for the limit dichroism are between -1.0 and -1.3 for chain lengths between 60 and 1000 base-pairs, whereas values around -1.4 are observed at chain lengths greater than 1000 base-pairs. These data indicate that electric fields extend the contour of DNA strands at high chain lengths from a weakly bent to a more linear form. The variations of the limit dichroism observed for short fragments suggest sequence-dependent differences in the secondary structure of the helix. The experimental results are compared with numerical calculations based on simple polyelectrolyte models. For short fragments the magnitude of several electrochemical parameters can be adequately explained by a polarization of the ion cloud around the DNA molecules. However, these polyelectrolyte models do not adequately describe the observed chain length dependence of the orientation phenomena.     

Endocrinological effects of strong 60-Hz electric fields on rats

Adult male rats were exposed or sham-exposed to 60-Hz electric fields without spark discharges, ozone, or significant levels of other secondary variables. No effects were observed on body weights or plasma hormone levels after 30 days of exposure at an effective field strength of 68 kV/m. After 120 days of exposure (effective field strength = 64 kV/m), effects were inconsistent, with significant reductions in body weight and plasma levels of follicle-stimulating hormone and corticosterone occurring in one replicate experiment but not in the other. Plasma testosterone levels were significantly reduced after 120 days of exposure in one experiment, with a similar but not statistically significant reduction in a replicate experiment. Weanling rats, exposed or sham-exposed in electric fields with an effective field strength of 80 kV/m from 20 to 56 days of age, exhibited identical or closely similar growth trends in body and organ weights. Hormone levels in exposed and sham-exposed groups were also similar. However, there was an apparent phase shift between the two groups in the cyclic variations of concentrations of hormones at different stages of development, particularly with respect to follicle-stimulating hormone and corticosterone. We concluded that 60-Hz electric fields may bring about subtle changes in the endocrine system of rats, and that these changes may be related to alterations in episodic rhythms.     

Electrical hemolysis of human and bovine red blood cells

Summary The external electric field strength required for electrical hemolysis of human red blood cells depends sensitively on the composition of the external medium. In isotonic NaCl und KCl solutions the onset of electrical hemolysis is observed at 4 kV per cm and 50% hemolysis at 6 kV per cm, whereas increasing concentrations of phosphate, sulphate, sucrose, inulin and EDTA shift the onset and the 50% hemolysis-value to higher field strengths. The most pronounced effect is observed for inulin and EDTA. In the presence of these substances the threshold value of the electric field strength is shifted to 14 kV per cm. This is in contrast to the dielectric breakdown voltage of human red blood cells which is unaltered by these substances and was measured to be 1 V corresponding in the electrolytical discharge chamber to an external electric field strength of 2 to 3 kV per cm. On the other hand, dielectric breakdown of bovine red blood cell membranes occurs in NaCl solution at 4 to 5 kV per cm and is coupled directly with hemoglobin release. The electrical hemolysis of cells of this species is unaffected by the above substances with exception of inulin. Inulin suppressed the electrical hemolysis up to 15 kV per cm. The data can be explained by the assumption that the reflection coefficients of the membranes of these two species to bivalent anions and uncharged molecules are field-dependent to a different extent. This explanation implies that electrical hemolysis is a secondary process of osmotic nature induced by the reversible permeability change of the membrane (dielectric breakdown) in response to an electric field. This view is supported by the observation that the mean volumes of ghost cells obtained by electrical hemolysis can be changed by changing the external phosphate concentration during hemolysis and resealing, or by subjecting the cells to a transient osmotic stress immediately after the electrical hemolysis step. An interesting finding is that the breakdown voltage, although constant throughout each normally distributed ghost size distribution, increases with increasing mean volume of the ghost populations.     

紫膜碎片的电二色性研究

悬浮在水中的嗜盐菌紫膜碎片,在外电场作用下产主定向排列.在20℃时,568nm的电二色性研究表明:外加电场为2kV/m时取向程度可达60%以上;大于5.5kV/m时,取向作用趋于饱和状态;饱和时简约电二色性为-0.437左右,视黄醛生色团的跃迁矩方向与电偶极矩方向形成60.9°夹角;紫膜的永久偶极短为9.2×10~(-24)C、M,剩余电极化率为3.0×10~(-27)m~2;紫膜的旋转扩散常数为0.53秒~(-1).曲线拟合分析表明,感应偶极对紫膜碎片的定向的贡献应予考虑.本文对紫膜碎片的定向机理进行了讨论.     

Detection of 60-hertz vertical electric fields by rats

Rats were trained to press levers to indicate the presence or absence of 60-Hz vertical electric fields at intensities from 0 to 27 kV/m (rms). The probability of detecting the field increased as the strength of the field increased. The shape of the detection curve (psychometric function) for most subjects (Ss) was similar whether the discriminative stimulus was the electric field or a tone. Two protocols were used to estimate the minimum field intensity necessary to detect the field (Reiz Limen, RL). The RL was estimated to be 13.3 kV/m (rms) when using one protocol (the staircase method) and 7.9 kV/m (rms) when using another protocol (the method of constant stimuli).     

Dynamics of lysozyme and its hydration water under an electric field

The effects of a static electric field on the dynamics of lysozyme and its hydration water are investigated by means of incoherent quasi-elastic neutron scattering (QENS). Measurements were performed on lysozyme samples, hydrated respectively with heavy water (D ₂O) to capture the protein dynamics and with light water (H ₂O), to probe the dynamics of the hydration shell, in the temperature range from 210 < T < 260 K. The hydration fraction in both cases was about ～ 0.38 gram of water per gram of dry protein. The field strengths investigated were respectively 0 kV/mm and 2 kV/mm ( ～2 × 10 ⁶ V/m) for the protein hydrated with D ₂O and 0 kV and 1 kV/mm for the H ₂O-hydrated counterpart. While the overall internal protons dynamics of the protein appears to be unaffected by the application of an electric field up to 2 kV/mm, likely due to the stronger intra-molecular interactions, there is also no appreciable quantitative enhancement of the diffusive dynamics of the hydration water, as would be anticipated based on our recent observations in water confined in silica pores under field values of 2.5 kV/mm. This may be due to the difference in surface interactions between water and the two adsorption hosts (silica and protein), or to the existence of a critical threshold field value E _c ～2–3 kV/mm for increased molecular diffusion, for which electrical breakdown is a limitation for our sample.     

Electrical hemolysis of human and bovine red blood cells.

The external electric field strength required for electrical hemolysis of human red blood cells depends sensitively on the composition of the external medium. In isotonic NaCl und KCl solutions the onset of electrical hemolysis is observed at 4 kV per cm and 50 per cent hemolysis at 6 kV per cm, whereas increasing concentrations of phosphate, sulphate, sucrose, inulin and EDTA shift the onset and the 50 per cent hemolysis-value to higher field strengths. The most pronounced effect is observed for inulin and EDTA. In the presence of these substances the threshold value of the electric field strength is shifted to 14 kV per cm. This is in contrast to the dielectric breakdown voltage of human red blood cells which is unaltered by these substances and was measured to be approximately 1 V corresponding in the electrolytical discharge chamber to an external electric field strength of 2 to 3 kV per cm. On the other hand, dielectric breakdown of bovine red blood cell membranes occurs in NaCl solution at 4 to 5 kV per cm and is coupled directly with hemoglobin release. The electrical hemolysis of cells of this species is unaffected by the above substances with exception of inulin. Inulin suppressed the electrical hemolysis up to 15 kV per cm. The data can be explained by the assumption that the reflection coefficients of the membranes of these two species to bivalent anions and uncharged molecules are field-dependent to a different extent. This explanation implies that electrical hemolysis is a secondary process of osmotic nature induced by the reversible permeability change of the membrane (dielectric breakdown) in response to an electric field. This view is supported by the observation that the mean volumes of ghost cells obtained by electrical hemolysis can be changed by changing the external phosphate concentration during hemolysis and resealing, or by subjecting the cells to a transient osmotic stress immediately after the electrical hemolysis step. An interesting finding is that the breakdown voltage, although constant throughout each normally distributed ghost size distribution, increases with increasing mean volume of the ghost populations.     

Occupational exposure to electric and magnetic fields during work tasks at 110 kV substations in the Tampere region

The occupational exposure to electric and magnetic fields during various work tasks at seven 110 kV substations in Finland's Tampere region was studied. The aim was to investigate if the action values (10 kV/m for the E‐field and 500 µT for the B‐field) of the EU Directive 2004/40/EC were exceeded. Electric and magnetic fields were measured during the following work tasks: (1) walking or operating devices on the ground; (2) working from a service platform; (3) working around the power transformer on the ground or using a ladder; and (4) changing a bulb from a man hoist. In work task 2 “working from a service platform” the measured electric field (maximum value 16.6 kV/m) exceeded 10 kV/m in three cases. In the future it is important to study if the limit value (10 mA/m²) of Directive 2004/40/EC is exceeded at 110 kV substations. The occupational 500 µT action value of the magnetic flux density field (B‐field) was not exceeded in any working situation. Bioelectromagnetics 31:252–254, 2010. © 2009 Wiley‐Liss, Inc.     

Perception and the Characteristics of the Response of Honey Bees,Paper Wasps,and Red Wood Ants to a Low-Frequency Electric Field

Bees, wasps, and ants have no specialized receptors for the perception of an electric field. An appropriate response to naturally occurring electric fields in bees and ants is associated with atmospheric exposure, amplified by the approach of the front of a thunderstorm. The primary transducers of mechanoreceptors that respond to displacement are related to the perception of low-frequency electric fields of high intensity by insects. The non-specific mechanism of perception of electric fields is based on irritation by induced currents that flow in the locations of their contact with each other and/or conductive surfaces. The frequency dependence of the electric field sensitivity is determined mainly by the magnitude of the current induced by it and the intensity of its contact action. The magnitude of the current induced in the outer part of the insect body is non-linearly related to the frequency of the electric field. The region with the highest sensitivity to electric fields is close to 500 Hz, which is consistent with the maximum magnitude of the induced current. At the same time, the threshold of the sensitivity to an electric field in wasps is approximately 0.04 kV/m, while in bees it is 0.45 kV/m. Ants react to the action of an electric field of 7–10 kV/m by slowing their movement. Magnetic fields and ionization, which accompany the generation of an electric field whose intensity reaches 15–20 kV/m, do not stimulate changes in the behavior of insects.

     

高压静电场对绵羊精子存活率的影响

采用不同剂量的高压静电场处理绵羊精液,经分析发现,高压静电场对绵羊精具有激活作用。能提高绵羊精液品质,表现在适当剂量的高压静电场能显著地提高绵羊精子存活率,其中以６００ｋＶ／ｍ剂量处理效果最佳。１００ｋＶ／ｍ和３００ｋＶ／ｍ剂量对精子刺激不足,而９００ｋＶ／ｍ剂量则对精子刺激过程,导致部分精子损伤和死亡,同样达不到预期的效果。     

Evaluation of the electrostatic field strength at the site of exocytosis in adrenal chromaffin cells.

Exocytosis in secretory cells consists of release from intracellular storage granules directly into the extracellular space via fusion of the granule membrane with the plasma membrane of the cell. It is considered here as comprising two distinct processes. One is the close apposition of granule and plasma membranes. The other arises from interactions between the two membranes during the process of apposition, leading to the formation of a fusion pore. In the following it is shown for the case of the adrenal medullary chromaffin cell that the fusion pore can be ascribed to electroporation of the granule membrane, triggered by the strong electric field existing at the site of exocytosis. Based on an electric surface charge model of the cytoplasmic side of the plasma membrane, resulting from the negatively charged phosphatidylserine groups, it is found that the electrostatic field strength at the site of exocytosis reaches values on the order of 10(8) V/m at small intermembrane distances of 3 nm and lower. The field strength increases with the size of the disc-shaped plasma membrane region generating the electric field, reaching an approximate limit for a radius of 10 nm, at a surface charge density of 5.4 x 10(-2) C/m2. According to previous experimental evaluations of threshold field strength, this field is sufficiently strong to cause membrane electroporation. This step is a precondition for the subsequent membrane fusion during the ongoing process of apposition, leading to secretion.     

Comparison of the coupling of grounded humans,swine and rats to vertical, 60-Hz electric fields

Published and new data for grounded humans, swine, and rats exposed to vertical, 60-Hz electric fields are used to determine field strengths at the surfaces of the bodies and average components of induced-current density along the axes of the bodies. At the tops of the bodies, surface electric fields are increased (enhanced) over the unperturbed field strength present before the subjects entered the field by factors of 17,7, and 4 for humans, swine, and rats, respectively. For an unperturbed field strength of 10 kV/m, average induced axial current densities in the neck, chest, abdomen, and feet are: 550, 190, 250, and 2000 nA/cm², respectively, for humans; 40, 13, 20, and 1100 nA/cm², respectively, for swine; and 28, 16, 2, and 1400 nA/cm², respectively, for rats. These data are used to show that the actual electric fields experienced by animals depend strongly on the shape of the body and its orientation relative to the electric field and ground plane. This fact must be taken into account if biological data obtained with laboratory animals are to be used for the assessment of possible hazards to humans exposed to 60-Hz electric fields.     

60-Hz electric field alters the steroidogenic response of rat adrenal tissue, in vitro

Exposure to a 60-Hz electric field at 10 kV/m but not at 5 kV/m, 100 kV/m or 1000 kV/m caused a highly significant, threefold elevation in the steroidogenic response of rat adrenal cortical tissue after the administration of 10 mU of adrenocorticotrophic hormone (ACTH) under in vitro, superfusion conditions. A 60-Hz electric field can directly influence the function of mammalian tissue in the absence of central-nervous-system mediation.     

Dose response study of human exposure to 60 Hz electric and magnetic fields

This human exposure study examined the relationship between field strength and biological response and tested whether the exposure levels at which the greatest effects occur differ for different endpoints. Three matched groups of 18 men each participated in two 6 h exposure test sessions. All subjects were sham exposed in one session. In the other session, each group of subjects was exposed at a different level of combined electric and magnetic field strength (low group: 6 kV/m, 10 μT; medium group: 9 kV/m, 20 μT; and high group: 12 kV/m, 30 μT). The study was performed double blind, with exposure order counterbalanced. Significant slowing of heart rate, as well as alterations in the latency and amplitude of event-related brain potential measures derived from the electro encephalogram (EEG), occurred in the group exposed to the 9 kV/m, 20 μT combined field (medium group). Exposure at the other field strength levels had no influence on cardiac measures and differential effects on EEG activity. Significant decrements in reaction time and in performance accuracy on a time estimation task were observed only in the low group. These results provide support for the hypothesis that humans may be more responsive to some combinations or levels of field strength than to others and that such differences in responsivity may depend, in part, on the endpoint of interest. © 1994 Wiley-Liss, Inc.     

The influence of electric field exposure on bone growth and fracture repair in rats

Rats were exposed to a 60-Hz electric field at an unperturbed field strength of 100 kV/m to determine its affect on bone growth and fracture repair. Exposure of immature male and female rats for 20 h/day for 30 days did not alter growth rate, cortical bone area, or medullary cavity area of the tibia. In another experiment, midfibular osteotomies were performed and the juvenile rats were exposed at 100 kV/m for 14 days. Evaluation by resistance to deformation and breaking strength indicated that fracture repair was not as advanced in the exposed animals as in the sham-exposed animals. In another experiment measurements of resistance to deformation were made in adult rats at 16, 20, and 26 days after osteotomy. Fracture repair was slower in exposed compared to control animals at day 20 and, to a lesser extent, at day 16, but not at day 26.     

60-Hz electric-field effects on pineal melatonin rhythms: time course for onset and recovery

Rats exposed for 3 weeks to uniform 60-Hz electric fields of 39 kV/m (effective field strength) failed to show normal pineal gland circadian rhythms in serotonin N-acetyl transferase activity and melatonin concentrations. The time required for recovery of the melatonin rhythm after cessation of field exposure was determined to be less than 3 days. The rapid recovery suggests that the overall metabolic competence of the pineal is not permanently compromised by electric-field exposure, and that the circadian rhythm effect may be neuronally mediated.     

Occupational exposure to electric fields and currents associated with 110 kV substation tasks

The main aim of this study was to investigate occupational exposure to electric fields, and current densities and contact currents associated with tasks at air-insulated 110 kV substations and analyze if the action value of EU Directive 2004/40/EC was exceeded. Four workers volunteered to simulate the following tasks: Task (A) maintenance of an operating device of a disconnector at ground or floor level, Task (B) maintenance of an operating device of a circuit breaker at ground or floor level, Task (C) breaker head maintenance from a man hoist, and Task (D) maintenance of an operating device of a circuit breaker from a service platform. The highest maximum average current density in the neck was 1.8 mA/m(2) (calculated internal electric field 9.0-18.0 mV/m) and the highest contact current was 79.4 μA. All measured values at substations were lower than the limit value (10 mA/m(2)) of the EU Directive 2004/40/EC and the 2010 basic restrictions (0.1 and 0.8 V/m for central nervous system tissues of the head, and all tissues of the head and body, respectively) of the International Commission on Non-Ionizing Radiation Protection (ICNIRP).