Thermal effects of 2.45 GHz microwaves on survival and viability of Chinese hamster V-79 cells

The possible existence of thermal effects specific to microwaves at 2.45 GHz and not found with classical heating in a waterbath was studied by measuring cell survival (colony-forming ability) and cell viability (the ability to exclude trypan blue) in Chinese hamster V79 cells. The microwaves were employed at high power densities (125 to 175 mW/cm2) corresponding to specific absorption rates ranging between 62 and 87 mW/g. When matching the rises in temperature, the effects of microwave-induced hyperthermia at 125 mW/cm2 on cell survival were comparable to those of classical heating. However, they were statistically significantly different when using power densities of 150 and 175 mW/cm2. The response obtained in terms of cell viability appeared to be comparable. The conclusions are also valid when taking into account a correction factor for energy losses during microwave treatment. The apparent specific effect of microwaves appears to be associated with exposures at high power densities involving short treatment times and rapid rises in temperature.     

Thermal aspects of biological effects of microwaves in Saccharomyces cerevisiae

The formation of zygotes between two haploid strains of yeast (Saccharomyces cerevisiae) was determined under treatment with microwaves of 9.4 and 17 GHz at power levels up to 50 and 60 mW/cm2 and a specific absorption rate below 24 mW/g, or with conventional heating. Microwave treatments at 9.4 GHz or 17 GHz at a power density of 10 mW/cm2 produced an increase in zygote formation equivalent to that produced by conventional heating in an incubator, i.e. equivalent to a rise in temperature of 0.5 or 1 degrees C. At higher power densities zygote formation was slightly increased by microwaves at 17 GHz as compared to microwaves at 9.4 GHz probably due to the higher absorption of microwaves at 17 GHz by intracellular water molecules. Under these conditions, microwaves had no effect on cell survival or the induction of cytoplasmic 'petite' mutations.     

Microwave radiation (2450 MHz) alters the endotoxin-induced hypothermic response of rats

The parenteral administration of bacterial endotoxin to rats causes a hypothermia that is maximal after approximately 90 minutes. When endotoxin-injected rats were held in a controlled environment at 22°C and 50% relative humidity and exposed for 90 minutes to microwaves (2450 MHz, CW) at 1 mW/cm², significant increases were observed in body temperature compared with endotoxintreated, sham-irradiated rats. The magnitude of the response was related to power density (10 mW/cm² > 5 mW/cm² > 1 mW/cm²). Saline-injected rats exposed for 90 minutes at 5 mW/cm² (specific absorption rate approximately 1.0 mW/g) showed no significant increase in body temperature compared with saline-injected, sham-irradiated rats. The hypothermia induced by endotoxin in rats was also found to be affected by ambient temperature alone. Increases in ambient temperature above 22°C in the absence of microwaves caused a concomitant increase in body temperature. This study reveals that subtle microwave heating is detectable in endotoxin-treated rats that have an impaired thermoregulatory capability. These results indicate that the interpretation of microwave-induced biological effects observed in animals at comparable rates and levels of energy absorption should include a consideration of the thermogenic potential of microwaves.     

Effects of 2.45-GHz microwave radiation on embryonic quail hearts

Although exposure to nonionizing electromagnetic radiation has been reported to cause a variety of systemic alterations during embryonic development, there are few reports of the induction of specific physiologic or morphologic changes in the myocardium. This study was designed to examine the effects of microwave radiation on cardiogenesis in Japanese quail embryos exposed during the first eight days of development to 2.45-GHz continuous-wave microwaves at power densities of 5 or 20 mW/cm². The specific absorption rates were 4.0 and 16.2 mW/g, respectively. The ambient temperature for each exposure was set to maintain the embryonated eggs at 37.5 °C. This did not preclude thermal gradients in the irradiated embryos since microwaves may not be uniformly absorbed. The test exposure levels did not induce changes in either the morphology of the embryonic heart or the ultrastructure of the myocardial cells. Analysis of the enzymatic activities of lactate dehydrogenase, glutamic oxaloacetic transaminase, and creatine phosphokinase failed to reveal any statistically significant differences between the nonexposed controls and those groups exposed to either 5 or 20 mW/cm². The data indicate that 2.45-GHz microwave radiation at 5 or 20 mW/cm² has no effect on the measured variables of the Japanese quail myocardium exposed during the first eight days of development.     

Microwave irradiation and in vitro release of enzymes from hepatic lysosomes

Summary An exposure system for examining in vitro effects of microwave irradiation on cellular and subcellar components has been developed. The system was used to test the effect of 2.45-GHz CW microwaves on the release of two lysosomal enzymes. At a specific absorption rate (SAR) of 10, 50 or 100 mW/g (90 min) no effects were noted at 37° C (pH of 7.3) on lysosomal fragility as determined by the release of the lysosomal enzymes cathepsin D and-glucuronidase. When the medium was adjusted to pH 5.0, microwave irradiation of the lysosomal suspension had no effect on the acid-induced enhancement of release of lysosomal enzymes. The data indicate that microwave irradiation had no labilizing effect on the lysosomal membrane, although other microwave-membrane interactions not associated with enzyme release may occur.     

Thermoregulatory physiologic responses in the human body exposed to microwave radiation

By introduction of an additional compartment in the hypothalamic region Stolwijk's thermo-regulatory model has been modified to consider partial heating due to hot spots induced by microwaves. It was found that because of thermoregulatory action, the temperature of the hypothalamus will not increase drastically until the rate of energy deposition exceeds the threshold level of about 50 mW/g. The primary controlling mechanisms are blood flow and sweating. For an energy deposition rate of 10 mW/g in the hypothalamus the increase in blood flow in the skin is negligible and the temperature rise of the hypothalamus as compared with blood temperature is about 0.5 °C. It was found that exposure of the head to electromagnetic radiation, in general, causes a decrease in temperature of the trunk and skin. The results show that while the deposition of energy in the hypothalamus at the rate of 10 mW/g produced significant conductive and convective effects, the same total energy uniformly distributed over the cranial cavity produces less significant effects.     

Thermoregulatory adjustments in squirrel monkeys exposed to microwaves at high power densities

The present study was undertaken to investigate the thermal adjustments of squirrel monkeys exposed in a cold environment to relatively high energy levels of microwave fields. The animals (Saimiri sciureus) were equilibrated for 90 min to a cool environment (Ta = 20 degrees C) to elevate metabolic heat production (M). They were then exposed for brief (10-min) or long (30-min) periods to 2,450-MHz continuous-wave microwaves. Power densities (MPD) were 10, 14, 19, and 25 mW/cm2 during brief exposures and 30, 35, 40, and 45 mW/cm2 during long exposures (rate of energy absorption: SAR = 0.15 [W/kg]/[mW/cm2]). Individual exposures were separated by enough time to allow physiological variables to return to baseline levels. The results confirm that each microwave exposure induced a rapid decrease in M. In a 20 degree C environment, the power density of a 10-min exposure required to lower M to approximate the resting level was 35 mW/cm2 (SAR = 5.3 W/kg). During the long exposures, 20 min was needed to decrease M to its lowest level. Cessation of irradiation was associated with persistence of low levels of M for periods that depended on the power density of the preceding microwave exposure. Vasodilation, as indexed by changes in local skin temperature, occurred at a high rate of energy absorption (SAR = 4.5 W/kg) and was sufficient to prevent a dramatic increase in storage of thermal energy by the body; vasoconstriction was reinstated after termination of irradiation. Patterns of thermophysiological responses confirm the influence both of peripheral and of internal inputs to thermoregulation in squirrel monkeys exposed to microwaves in a cool environment.     

Effect of microwaves (2450-MHz) on the immune system in mice: studies of nucleic acid and protein synthesis

CBA/J adult male mice were given single or triple exposures to 2450-mHz microwaves in an environmentally controlled wave guide facility. The average absorbed dose rate for a single exposure varied from 12 to 15 mW/g. Sham-exposed mice served as controls. Lymphoid cells were collected and tested for metabolic activity on days 3, 6, and 9 following a single exposure, and on days 9, 12, and 16 following triple exposures on days 0, 3, and 6. Cells were cultured in vitro for four hours to seven days before their metabolic rates were assayed. Under these conditions, microwaves failed to produce any detectable change in deoxyribonucleic acid (DNA), ribonucleic acid (RNA), and protein synthesis, as measured by the incorporation of methyl(3H)-thymidine (3H-TDR) (DNA substrate), 3H-uridine (3H-UR) (RNA substrate), and 3H-leucine (protein substrate) by spleen, bone marrow, and peripheral blood lymphocytes (PBL) in vitro. These data suggest that microwave-induced increases in the frequency of complement-receptor (CR)- or surface-immunoglobulin (sIg)-bearing cells were not associated with a concomitant increase in cell proliferation and/or protein synthesis, and favor the concept that microwaves under these conditions stimulate already existing B-cell precursors for maturation.     

Comparison of microwaves to fluidized sand baths for heating tubular reactors for hydrothermal and dilute acid batch pretreatment of corn stover

Heating of batch tubular reactors with fluidized sand baths and with microwaves resulted in distinctive sugar yield profiles from pretreatment and subsequent enzymatic hydrolysis of corn stover at the same time, temperature, and dilute sulfuric acid concentration combinations and hydrothermal pretreatment conditions. Microwave heated pretreatment led to faster xylan, lignin, and acetyl removal as well as earlier xylan degradation than sand baths, but maximum sugar recoveries were similar. Solid state CP/MAS NMR revealed that microwave heating was more effective in altering cellulose structural features especially in breakdown of amorphous regions of corn stover than sand bath heating. Enzymatic hydrolysis of pretreated corn stover was improved by microwave heating compared to sand bath heating. Mechanisms were proposed to explain the differences in results for the two systems and provide new insights into pretreatment that can help advance this technology.     

Quantification of metabolic activity of cultured plant cells by vital staining with fluorescein diacetate

The metabolic activity of suspension cultures of Sonneratia alba cells was quantified by measurement of the hydrolysis of fluorescein diacetate (FDA). FDA is incorporated into live cells and is converted into fluorescein by cellular hydrolysis. Aliquots (0.1–0.75 g) of S. alba cells were incubated with FDA at a final concentration of 222 μg/ml suspension for 60 min. Hydrolysis was stopped, and fluorescein was extracted by the addition of acetone and quantified by measurement of absorbance at 490 nm. Fluorescein was produced linearly with time and cell weight. Cells of S. alba are halophilic and proliferated well in medium containing 50 and 100 mM NaCl. Cells grown in medium containing 100 mM NaCl showed 2- to 3-fold higher FDA hydrolysis activity than those grown in NaCl-free medium. When S. alba cells grown in medium supplemented with 50 mM NaCl were transferred to fresh medium containing 100 mM mannitol, cellular FDA hydrolysis activity was down-regulated after 4 days of culture, indicating that the moderately halophilic S. alba cells were sensitive to osmotic stress. Quantification of cellular metabolic activity via the in vivo FDA hydrolysis assay provides a simple and rapid method for the determination of cellular activity under differing culture conditions.     

Millimeter microwave effect on ion transport across lipid bilayer membranes

The effects of millimeter microwaves in the frequency range of 54–76 GHz on capacitance and conductance of lipid bilayer membranes (BLM) were studied. Some of the membranes were modified by gramicidin A and amphotericin B or by tetraphenylboron anions (TPhB_?). The millimeter microwaves were pulse-modulated (PW) at repetition rates ranging from 1 to 100 pps, PW at 1000 pps, or unmodulated continuous waves (CW). The maximum output power at the waveguide outlet was 20 mW. It was found that CW irradiation decreased the unmodified BLM capacitance by 1.2% ± 0.5%. At the same time, membrane current induced by TPhB transport increased by 5% ± 1%. The changes in conductance of ionic channels formed by gramicidin A and amphotericin B were small (0.6% ± 0.4%). No “resonance-like” effects of mm-wave irradiation on membrane capacitance, ionic channel currents, or TPhB transport were detected. All changes in membrane capacitance and currents were independent of the modulation employed and were equivalent to heating by approximately 1.1 °C. © 1995 Wiley-Liss, Inc.     

Noise-modulated-microwave-induced response in snail neurons

Helix aspersa neurons were irradiated with noise-amplitude-modulated microwaves (carrier frequency 2450 MHz, 20% AM, 0-20 kHz, specific absorption rate 6.8 and 14.4 mW/g). It was found that such an exposure caused an appearance of high frequency bursts and a rise in membrane resistance.     

Effect of 9.6-GHz pulsed microwaves on the orb web spinning ability of the cross spider (Araneus diadematus)

Eight cross spiders (Araneus diadematus) were exposed overnight (16 h) during web-building activity to pulsed 9.6-GHz microwaves at average power densities of 10, 1, and 0.1 mW/cm2 (estimated SARs 40, 4, and 0.4 mW/g). Under these conditions, 9.6-GHz pulsed microwaves did not affect the web-spinning ability of the cross spider.     

Operant control of convective cooling and microwave irradiation by the squirrel monkey

Adult male squirrel monkeys (Saimiri sciureus) were individually chair-restrained in an air-conditioned Styrofoam box in the far field of a horn antenna. Each monkey first received extensive training to regulate the temperature of the air circulating through the box by selecting between 10 and 50 degrees C air source temperatures. Then, to investigate the ability of the animals to utilize microwaves as a source of thermalizing energy, 2450-MHz continuous wave microwaves accompanied by thermoneutral (30 degrees C) air were substituted for the 50 degrees C air. Irradiation at each of three power densities was made available, ie, at 20, 25, and 30 mW/cm2 [SAR = 0.15 (W/kg)/(mW/cm2)]. The percentage of time that the monkeys selected microwave irradiation paired with thermoneutral air averaged 90% at 20 and at 25 mW/cm2. The mean percentage declined reliably (p less than 0.001) to 81% at 30 mW/cm2, confirming the monkey's ability to utilize microwave irradiation as a source of thermal energy during the course of behavioral thermoregulation. All animals readily made the warm-air to microwave-field transition, regulating rectal temperature with precision by sequentially selecting 10 degrees C air, then microwave irradiation accompanied by 30 degrees C air. Although the selection of cooler air resulted in a slight reduction of skin temperatures, normal rectal temperature was maintained. The results indicate that the squirrel monkey can utilize a microwave source in conjunction with convective cooling to regulate body temperature behaviorally.     

Microwave esterification of sunflower proteins in solvent-free conditions

The esterification of storage proteins from sunflower seeds renders these molecules more hydrophobic and increases internal plasticization, facilitating their use in the fabrication of materials by thermo-mechanical processes. The reaction was carried out in solvent-free conditions to simplify the process and to reduce costs. Optimization of esterification by the classical method, heating in a thermostat-regulated oil bath, resulted in 84% esterification for a reaction time of 4 h (amount of catalyst = 3.9 meq 5 N HCl/g protein, T = 90 degrees C). Microwave heating was then used to reduce the reaction time and the hydrolysis of protein chains. A similar level of esterification (89%) was obtained in 18 min (amount of catalyst = 3.4 meq 5 N HCl/g protein, microwave power (P) = 560 W). High yields were obtained with this method, demonstrating that this process limited hydrolysis.     

Microwave absorption by magnetic nanoparticles in the organism

An estimate is presented for the rate of microwave absorption by magnetic nanoparticles in the organism. The absorption takes place because of energy dissipation at ferromagnetic resonance. Based on the known solution of the Landau-Lifshits equation, the imaginary part of complex susceptibility is evaluated, which gives the absorption rate. It is shown that even under thermal isolation of the particles, their heating would be insignificant upon absorption of radiation with an energy flux density on the order of 1 mW/cm², and such a mechanism is unrelated to the observable effects of low-intensity microwaves.     

The role of TLR4 receptor in the stress response of lymphocytes

In vitro effects of low-level electromagnetic waves (8.18 GHz, frequency swings within 1 s, intensity 1 microW/cm, exposure for 1 h) and low-energy laser light (He-Ne laser with 632.8 nm, 0.2 mW/cm, dose 1.2 x 10(-2) J/cm2) on the expression of receptor protein TLR4, which is known as a part of the system for microbal toxin recognition, were studied in mouse lymphocytes. In addition, TLR4 expression was examined in situations when stress responses to low-level nonionizing radiation were modified by the antibiotic geldanamycin, which suppresses the activity of the heat shock protein Hsp90. It was found that low-level microwaves significantly raised the amount of TLR4; in contrast, laser light decreased the expression of the receptor in lymphocytes. In cells pretreated with geldanamycin, the TLR4 expression in irradiated cells was reduced to minimum levels, much lower than control values. The results showed that TLR4, which is involved in specific binding of toxin from gram-negative bacteria, can regulate cell responses to signals of other origin, in particular to nonionizig radiation, including low-level microwaves and laser light.     

Analysis of enzyme kinetics in individual living cells utilizing fluorescence intensity and polarization measurements

BACKGROUND: The Cellscan mark-S (CS-S) scanning cytometer was used for tracing enzymatic reactions in the same individual cells under various physiological conditions over periods of minutes. On-line reagent addition and changes in the experimental conditions (buffers, ions, substrates and inhibitors) were performed. METHODS: Kinetic events were monitored by fluorescence intensity (FI) and fluorescence polarization (FP) measurements of fluorescein diacetate (FDA) and chloromethyl fluorescein diacetate (CMFDA) intracellular hydrolysis. FP measurements have been used to assess the intracellular marker's mobility restrictions. RESULTS: Kinetic measurement along 1000 s of FDA labeled individual Jurkat T cells, indicated variation of 65% for FI(t) and approximately 10% for FP(t). While FI increased linearly with time, FP(t) decreased nonlinearly and asymptotically, reaching a constant value. The FP(t) of CMFDA-labeled cells was different from that of FDA-labeled cells. Average cellular Km of 3.9 microM was calculated from individual cell FDA hydrolysis curves. CONCLUSIONS: (1) Analysis of the reaction kinetics of intracellular enzymes can be refined by using FP measurements of the products of fluorogenic substrates in addition to the FI measurements. (2) Subpopulations or individual cells could be classified according to their reaction rates. (3) A specific dependence of FP(t) on type of enzyme substrate is suggested.     

Thermoregulatory, metabolic, and cardiovascular response of rats to microwaves.

This study was undertaken to determine the effects of 2,450-MHz microwave irradiation on thermoregulation, metabolism, and cardiovascular function of rats. Young adult male animals (430 g) were exposed for 30 min to 2,450-MHz microwaves in a cavity at absorbed dose rates of 0, 4.5, 6.5, or 11.1 mW/G. For animals of the size used in this study, these dose rates represent absorption of energy at the rate of 27.7, 40.1, and 68.2 cal/min, respectively. For a period of 5 h following exposure, measurements were made of colonic temperature, skin temperature, oxygen consumption, carbon dioxide production, respiratory quotient, and heart rate. Rats that received 27.7 cal/min for 30 min exhibited an initial transient increase in colonic and skin temperatures but no alterations in other functions. The group irradiated at 40.1 cal/min had greater elevations in colonic and skin temperatures immediately after exposure, followed by overcompensation and lower than normal colonic temperatures for about 3 h. The metabolic rate was depressed in this group for 3 h. Bradycardia developed within 20 min after exposure and persisted for about 3 h. The group of rats that received 68.2 cal/min for 30 min had responses similar to those of the 40.1 cal/min group, but the changes were more severe and lasted longer. In addition, a number of transient abnormalities were noted in the ECG tracings of rats that had received the highest dose, including irregular rhythms and incomplete heart block. The physiological changes observed in this study can be attributed to the heating induced by irradiation.