Long-term biological effects of air ions and D.C. electric fields on Namru mice: Second year report

This report describes the second year of long-term continuous exposures of female NAMRU mice to small air ions and D.C. electric fields in the following conditions: ± high ions ((2×10⁵/cm³), ± low ions (2×10³/cm³), ± field (2 kV/m) only and ground (ion depleted, no field). Using an isolated anesthesized mouse, whole body ion flux values averaged 1.04±0.63×10^–10 A in high ion cages for different positions on the cage floor, with about a hundred-fold reduction for low ion cages.During the second year (sample periods 5–8) of exposure serum chemistry variability increased, due to increased pathology and decreased numbers of animals as our experimental population died off. The fifth sample period yielded results consistent with those seen earlier, but later sample periods had many fewer significant differences between cages than did those of the first year. Nevertheless, MCA statistics for serum glucose for the second year found a pattern remarkably similar to the first, with the low ion cages (LN and LP) having the lowest levels. MCA statistics for both years emphasized this possible window effect of low level ionized conditions. Also, a comparison between the combined values for ionized (HN, LN, HP and LP) and ion depleted cages (NF, PF, G₁ and G₂) showed a highly significant difference (p<10^–6) for serum glucose for both years of exposure, with lower glucose values seen for animals in the ionized cages overall. Animals of all conditions also showed a highly significant decrease in serum glucose with age.Comparison of mice in ionized cages vs. the non-ionized cages also resulted in a significant difference (p<.013) for survival characteristics between groups, with ion exposed animals having a shorter lifespan. These statistics argue strongly for significant effects of long-term exposure of NAMRU mice to the ionized environment.     

Electric fields,small air ions and biological effects

Early growth of barley seedlings supplied with ion-depleted air in a controlled microenvironment was used as the criterion of physiological action in experiments designed to separate electrical field and air ion effects. The essential element in this work was the fact that³H foils, functioning as bias electrodes at applied voltages of 67.5-955 V and providing field strengths of 4.6 V/cm to 90 V/cm will produce 2.7×10⁴–1.7×10⁵ small air ions/cm³ — concentrations sufficient to yield a statistically valid increase in growth rate. In contrast, seedlings exposed to equivalent electrical fields in ion-depleted air exhibited no increase in rate of growth. These results indicate that this particular physiological response is produced by small negative air ions and is not dependent on the accompanying electrical fields.     

Behavioral monitoring of rats during exposure to air ions and DC electric fields

Air ions and direct current (DC) electric fields have been reported to exert subtle behavioral and biological effects on rodents and humans. These effects often appear inconsistent, yet there have been few attempts to resolve these inconsistencies by experimental replication. Rats exposed to negatively or positively charged air ions over a wide range of concentrations and exposure periods have been reported to show alterations in their level of locomotor activity. In this study, locomotor activity of Sprague-Dawley rats was quantified during exposure to either unipolar air ions and DC fields of the same polarity or DC fields alone. Both polarities were studied. Air ion concentrations were 5.0 X 10(3), DC fields were 3 kV/m, and exposures lasted 2, 18, or 66 h. In one experiment rats were exposed to DC fields of 12 kV/m. No exposure condition exerted any effect on locomotor activity or rearing behavior. In addition, no behavioral perturbations were observed after the onset of any of the exposure conditions, suggesting that the rats may have failed to detect the altered environment.     

Failure to produce taste-aversion learning in rats exposed to static electric fields and air ions

Taste-aversion (TA) learning was measured to determine whether exposure to high-voltage direct current (HVdc) static electric fields can produce TA learning in male Long Evans rats. Fifty-six rats were randomly distributed into four groups of 14 rats each. All rats were placed on a 20 min/day drinking schedule for 12 consecutive days prior to receiving five conditioning trials. During the conditioning trials, access to 0.1 % sodium saccharin-flavored water was given for 20 min, followed 30 min later by one of four treatments. Two groups of 14 rats each were individually exposed to static electric fields and air ions, one group to +75 kV/m (+2 × 10⁵ air ions/cm³) and the other group to ?75 kV/m (-2 × 10⁵ air ions/cm³). Two other groups of 14 rats each served as sham-exposed controls, with the following variation in one of the sham-exposed groups: This group was subdivided into two subsets of seven rats each, so that a positive control group could be included to validate the experimental design. The positive control group (n = 7) was injected with cyclophosphamide 25 mg/kg, i.p., 30 min after access to saccharin-flavored water on conditioning days, whereas the other subset of seven rats was similarly injected with an equivalent volume of saline. Access to saccharin-flavored water on conditioning days was followed by the treatments described above and was alternated daily with water “recovery” sessions in which the rats received access to water for 20 min in the home cage without further treatment. Following the last water-recovery session, a 20 min, two-bottle preference test (between water and saccharin-flavored water) was administered to each group. The positive control group did show TA learning, thus validating the experimental protocol. No saccharin-flavored water was consumed in the two-bottle preference test by the cyclophosphamide-injected, sham-exposed group compared to 74% consumed by the saline-injected sham-exposed controls (P <.0001). Saccharin-preference data for the static field-exposed groups showed no TA learning compared to data for sham-exposed controls. In summary, exposure to intense static electric fields and air ions did not produce TA learning as assessed by this particular design. © 1995 Wiley-Liss, Inc.     

Nanosecond pulsed electric fields (nsPEF) induce direct electric field effects and biological effects on human colon carcinoma cells

Nanosecond pulsed electric fields (nsPEFs) are ultrashort pulses with high electric field intensity (kV/cm) and high power (megawatts), but low energy density (mJ/cc). To determine roles for p53 in response to nsPEFs, HCT116 cells (p53+/+ and p53-/-) were exposed to nsPEF and analyzed for membrane integrity, phosphatidylserine externalization, caspase activation, and cell survival. Decreasing plasma membrane effects were observed in both HCT116p53+/+ and p53-/- cells with decreasing pulse durations and/or decreasing electric fields. However, addition of ethidium homodimer-1 and Annexin-V-FITC post-pulse demonstrated greater fluorescence in p53-/- versus p53+/+ cells, suggesting a postpulse p53-dependent biological effect at the plasma membrane. Caspase activity was significantly higher than nonpulsed cells only in the p53-/- cells. HCT116 cells exhibited greater survival in response to nsPEFs than HL-60 and Jurkat cells, but survival was more evident for HCT116p53+/+ cells than for HCT116p53-/- cells. These results indicate that nsPEF effects on HCT116 cells include (1) apparent direct electric field effects, (2) biological effects that are p53-dependent and p53-independent, (3) actions on mechanisms that originate at the plasma membranes and at intracellular structures, and (4) an apparent p53 protective effect. NsPEF applications provide a means to explore intracellular structures and functions that can reveal mechanisms in health and disease.     

The effects of air ions on brain levels of serotonin in mice

Mice were maintained in a controlled pollutant-free microenvironment and were exposed for 12, 24, 48 and 72 hr to 3 different concentrations of small positive or negative air ions: 2–4 × 10³ ions/cm³, 3–4 × 10⁴ ions/cm³ or 3.5–5 × 10⁵ ions/cm³. Spectrophotofluorometric assays of brain serotonin levels of air ion-treated mice showed statistically significant differences as early as 12 hours from those of mice kept in untreated pollutant-free air. Essentially no deviation from control values were observed at 24 and 48 hours. After 72 hours of exposure sharp decreases took place in all groups with the single exception of the animals exposed to 3–4 × 10⁴ positive ions/cm³. The hypothesis that alterations in mood and affect associated with certain meteorological conditions, e.g. winds such as the foehn, sirocco, etc. might depend upon air ion-induced changes in brain levels of serotonin was examined in the light of recent advances in neurophysiology and neuropharmacology.     

Exposure chamber for determining the biological effects of electric and magnetic fields on dairy cows

An exposure chamber was designed to study the effects of electric and magnetic fields (EMF) on oestrous cycles, hormonal profile during gestation, pineal function, quantity and quality of milk production, feed intake, and central nervous system of dairy cattle. The chamber was 15 x 10 x 3 m; and the control system was fully computerized so that the field intensities can be varied and monitored continuously, on site or remotely. During exposure to EMF, milk production, feed consumption, and health were monitored closely and blood and cerebral spinal fluid were continuously sampled. The chamber characteristics allow use of a wide range of exposure such as electric fields (0-30 kV/m) and magnetic fields (0-100 microT) at frequencies ranging from 45 to 3000 Hz.     

Long-term effects of 60-Hz electric vs. magnetic fields on IL-1 and IL-2 activity in sheep

This study was designed to assess the effect of exposure to long-term extremely low-frequency electric and magnetic fields (ELF-EMF) from a 500 kV transmission line on IL-1 and IL-2 activity in sheep. The primary hypothesis was that the reduction in IL-1 activity observed in our two previous short-term studies (10 months) was due to EMF exposure from this transmission line. To repeat and expand these studies and to characterize the components of EMF responsible for the previously observed reduction in IL-1 activity, the current experiment examined not only the effect of exposure to electric and magnetic fields, but also the magnetic field component alone. In the current study, IL-2 was examined to characterize the effects of EMF exposure on an indicator of T cell responses. 45 Suffolk ewe lambs were randomized into three groups of 15 animals each. One group of animals was placed in the EMF pen, located directly beneath the transmission line. A second group was placed in the shielded MF (magnetic field only) pen, also directly beneath the transmission line. The third group of animals was placed in the control pen located several hundred meters away from the transmission line. During the 27 month exposure period, blood samples were taken from all animals monthly. When the data were analyzed collectively over time, no significant differences between the groups were found for IL-1 or IL-2 activity. In previous studies ewe lambs of 8-10 weeks of age were used as the study animals and significant differences in IL-1 activity were observed after exposure of these animals to EMF at mean magnetic fields of 3.5-3.8 microT (35-38 mG) and mean electric fields of 5.2-5.8 kV/m. At the start of the current study EMF levels were reduced as compared to previous studies. One interpretation of the current data is that magnetic field strength and age of the animals may be important variables in determining whether EMF exposure will affect IL-1 activity.     

Alignment of microscopic particles in electric fields and its biological implications.

It is well known that electromagnetic fields cause mechanical forces. If one applies an electrical field to a suspension of microscopic particles, these particles realign themselves along the direction of the field and form pearl-chain-like aggregates. These chains are mostly single stranded but they are frequently multistranded. This phenomenon has been investigated by a number of groups. Here we discuss the dependence of threshold field strength on particle size and frequency. Also, pulsed fields have been thought to be more effective than continuous fields of the same average power in evoking biological effects. Our measurement of the threshold power requirement for the pearl-chain formation indicates that pulsed fields require as much power as continuous fields. The biological significance of pearl-chain formation is briefly discussed.     

The standardization of experimental investigations of biological effects of low frequency electric and magnetic fields

Many investigations show that electric and magnetic fields of low frequency may cause biological effects. However, the results of experiments differ considerably. In this paper, possible reasons for the limited reproducibility are pointed out, i.e., problems of generation, detection and definition of artificial fields. In particular, it is shown mathematically that the usual statements of electric field intensity are strongly misleading. The actual intensity is proportional to the ratio of the object height and the electrode distance as well as to the thinness of the object. Recommendations for the standardization of future investigations are given.     

A precision controlled environmental chamber for studies of the effects of electric fields on biological objects

An environmental chamber with associated control equipment for the study of possible effects of electric fields on biological objects (small mammals) is described in detail. Temperature is controlled within 0.4 C, relative humidity within ± 1.5 % and pressure below or above ambient is regulated to ± 1.0 mm Hg.Sound and electrical noise absorption by the chamber insulation is better than 90%.

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Zusammenfassung Eine Kammer mit dem zugehörigen KontrollgerÄt zum Studium der biologischen Wirkung von elektrischen Feldern an kleinen Versuchstieren wird ausführlich beschrieben.Die Temperatur ist innerhalb ±0,3C, die relative Feuchtigkeit innerhalb ± 1,5 % konstant und der Druck unterhalb oder oberhalb des Umgebungsdruckes auf ±1,0 mm Hg geregelt. Die LÄrmabsorption durch das Isolationsmaterial der Kammer ist über 90%.

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Resume On décrit ici en détails une chambre permettant l'étude des effets biologiques des champs électriques sur de petits mammifères. Cette chambre est assortie d'appareils de contrÔle. On peut y maintenir une température et une humidité constante (±0,3C, respectivement ±,1,5%) et la pression peut y Être réglée à volonté au-dessus et audessous de la pression ambiante avec une précision de ± 1,0 mm/Hg. L'absorption du bruit par le matériau d'isolation est supérieur à 90%.

     

Diverse effects of nanosecond pulsed electric fields on cells and tissues

The application of pulsed electric fields to cells is extended to include nonthermal pulses with shorter durations (10-300 ns), higher electric fields (< or =350 kV/cm), higher power (gigawatts), and distinct effects (nsPEF) compared to classical electroporation. Here we define effects and explore potential application for nsPEF in biology and medicine. As the pulse duration is decreased below the plasma membrane charging time constant, plasma membrane effects decrease and intracellular effects predominate. NsPEFs induced apoptosis and caspase activation that was calcium-dependent (Jurkat cells) and calcium-independent (HL-60 and Jurkat cells). In mouse B10-2 fibrosarcoma tumors, nsPEFs induced caspase activation and DNA fragmentation ex vivo, and reduced tumor size in vivo. With conditions below thresholds for classical electroporation and apoptosis, nsPEF induced calcium release from intracellular stores and subsequent calcium influx through store-operated channels in the plasma membrane that mimicked purinergic receptor-mediated calcium mobilization. When nsPEF were applied after classical electroporation pulses, GFP reporter gene expression was enhanced above that observed for classical electroporation. These findings indicate that nsPEF extend classical electroporation to include events that primarily affect intracellular structures and functions. Potential applications for nsPEF include inducing apoptosis in cells and tumors, probing signal transduction mechanisms that determine cell fate, and enhancing gene expression.     

Modeling environment for numerical simulation of applied electric fields on biological cells

The application of electric pulses in cells increases membrane permeability. This phenomenon is called electroporation. Current electroporation models do not explain all experimental findings: part of this problem is due to the limitations of numerical methods. The Equivalent Circuit Method (ECM) was developed in an attempt to solve electromagnetic problems in inhomogeneous and anisotropic media. ECM is based on modeling of the electrical transport properties of the medium by lumped circuit elements as capacitance, conductance, and current sources, representing the displacement, drift, and diffusion current, respectively. The purpose of the present study was to implement a 2-D cell Model Development Environment (MDE) of ionic transport process, local anisotropy around cell membranes, biological interfaces, and the dispersive behaviour of tissues. We present simulations of a single cell, skeletal muscle, and polygonal cell arrangement. Simulation of polygonal form indicates that the potential distribution depends on the geometrical form of cell. The results demonstrate the importance of the potential distributions in biological cells to provide strong evidences for the understanding of electroporation.     

First report of Montagnea arenaria (D.C.) Zeller (Agaricales) in Chile

Montagnea arenaria (D.C.) Zeller (Agaricales), an interesting basidiomycete adapted to arid environments is recorded for the first time from a sample collected on sandy soil in the northern zone of Chile. Its mature fruiting body is described. Taxonomic and ecological comments are also addressed.