Large scale in vitro experiment system for 2 GHz exposure

A beam formed radiofrequency (RF) exposure-incubator employing a horn antenna, a dielectric lens, and a culture case in an anechoic chamber is developed for large scale in vitro studies. The combination of an open type RF exposure source and a culture case through which RF is transmitted realizes a uniform electric field (+/-1.5 dB) in a 300 x 300 mm area that accommodates 49 35 mm diameter culture dishes. This large culture dish area enables simultaneous RF exposure of a large number of cells or various cell lines. The RF exposure source operates at 2142.5 MHz corresponding to the middle frequency of the downlink band of the International Mobile Telecommunication 2000 (IMT-2000) cellular system. The dielectric lens, which has a gain of 7 dB, focuses RF energy in the direction of the culture case and provides a uniform electric field. The culture case is sealed and connected to the main unit for environmental control, located outside the anechoic chamber, via ducts. The temperature at the center of the tray, which contains the culture dishes in the culture room, is maintained at 37.0 +/- 0.2 degrees C by air circulation. In addition, the appropriate CO2 density and humidity supplied to the culture case realizes stable long-term culture conditions. Specific absorption rate (SAR) dosimetry is performed using an electric field measurement technique and the Finite Difference Time Domain (FDTD) calculation method. The results indicate that the mean SAR of the culture fluid at the bottom of the 49 (7 x 7 array) culture dishes used in the in vitro experiments is 0.175 W/kg for an antenna input power of 1 W and the standard deviation of the SAR distribution is 59%. When only 25 culture dishes (5 x 5 array) are evaluated, the mean SAR is 0.139 W/kg for the same antenna input power and the standard deviation of the SAR distribution is 47%. The proliferation of the H4 cell line in 72 h in a pair of RF exposure-incubators reveals that the culture conditions are equivalent to those of a common CO2 incubator.     

Numerical modeling and dosimetry of the 35 mm Petri dish under 46 GHz millimeter wave exposure

Experimental studies on effects of millimeter wave (MMW) exposure on cells cultured in Petri dishes have attracted interest in recent decades. To improve the quantification of the biological responses toward the MMW energy, an accurate and precise MMW dosimetry is to be provided. By using the finite difference time domain (FDTD) method, the numerical dosimetry is performed for a typical 35 mm Petri dish under 46 GHz continuous MMW exposure from an irradiator of a specified power pattern. With the aim of building a precise model, the meniscus at the interface between the culture solution and the Petri dish sidewall is taken into account, followed by the modeling of smooth edges of the Petri dish. The trilinear interpolation is introduced to assist the FDTD method to obtain a more precise dosimetric assessment. The specific absorption rate (SAR) distributions in the cornea cells covered by culture solution in the Petri dish are calculated and compared to display the effects of using Petri dish models of various precision and the trilinear interpolation on dosimetry results. In addition, the SAR distribution in the cells is analyzed to study its homogeneity. The results indicate that the precise Petri dish model and the application of the trilinear interpolation are helpful in improving the precision of numerical dosimetry. It is also revealed that the inhomogeneity of the SAR distribution is well beyond neglect, which deserves cautious consideration in experiments investigating MMW effects on cells in vitro.     

Electromagnetic and thermal evaluation of an applicator specialized to permit high‐resolution non‐perturbing optical evaluation of cells being irradiated in the W‐band

To permit epi‐illuminated, high‐resolution optical microscopy of cells in monolayer culture during unperturbed W‐band (75–110 GHz) irradiation, a new class of applicator has been developed based upon WR10 rectangular waveguide components: the cells are normally plated onto the underside of a coverslip which is then placed against the under side of a waveguide flange and receives a roughly circular exposure pattern, with the ±1 dB central spot roughly 1 mm in diameter. Constructed and tested with 94 GHz millimeter waves, water‐immersion optics, and free‐convection cooling, the applicator works robustly and permits SARs at the cell layer as high as 4500 W/kg before the steady‐state temperature rise at the cell layer exceeds 0.5 K. Bioelectromagnetics 31:140–149, 2010. © 2009 Wiley‐Liss, Inc.     

Proposal of 28 GHz In Vitro Exposure System Based on Field Uniformity for Three‐Dimensional Cell Culture Experiments

This paper proposes a novel in vitro exposure system operating at millimeter‐wave (mmWave) 28 GHz, one of the frequency bands under consideration for fifth generation (5G) communication. We employed the field uniformity concept along cross‐sectional observation planes at shorter distances from the radiation antenna for better efficiency and a small‐size system. A choke‐ring antenna was designed for this purpose in consideration of a wider beamwidth (BW) and a symmetric far‐field pattern across three principal planes. The permittivity of Dulbecco's modified Eagle's medium solution was measured to examine the specific absorption rate (SAR) of the skin cell layer inside a Petri dish model for a three‐dimensional (3D) cell culture in vitro experiment. The best deployment of Petri dishes, taking into account a geometrical field symmetry, was proposed. Local SAR values within the cell layer among the Petri dishes were determined with different polarization angles. It was determined that this polarization effect should be considered when the actual exposure and deployment were conducted. We finally proposed an in vitro exposure system based on the field uniformity including downward exposure from an antenna for 3D cell culture experiments. A small‐size chamber system was obtained, and the size was estimated using the planar near‐field chamber design rule. Bioelectromagnetics. 2019;40:445–457. © 2019 Bioelectromagnetics Society     

Systems for exposing mice to 2,450-MHz electromagnetic fields

Two systems for exposing mice to 2,450-MHz electromagnetic fields are described. In a waveguide system, four mice were placed in a Styrofoam cage and exposed dorsally to circularly polarized electromagnetic fields. The temperature and humidity in the mouse holder were kept constant by forced-air ventilation. For 1-W input power to the waveguide, the average specific absorption rate (SAR) was determined by twin-well calorimetry to be 3.60 ± 0.11 (SE) W/kg in 27-g mice. The maximum SAR at the skin surface determined thermographically was 8.36 W/kg in the head of the mouse. The second system was a miniature anechoic chamber. Six mice were irradiated dorsally to far field plane waves. Copper shielding and high-temperature absorbing material were lined inside the chamber to accommodate the high input power. The air ventilation at the location of the mice was separately controlled so that any heating in the absorber would not affect the animals. For 1-W input power, the average SAR was 0.17 ± 0.01 W/kg and the maximum SAR at the skin surface was 0.41 W/kg in the animal when irradiated with body axis parallel to the E field; the SARs were 0.11 ± 0.01 W/kg and 0.64 W/kg, respectively, when irradiated perpendicular to the E field.     

Dosimetry evaluation of a cylindrical waveguide chamber for unrestrained small rodents at 1.9 GHz

An exposure system, consisting of four identical cylindrical waveguide chambers, was developed for studying the effects of radiofrequency (RF) energy on laboratory mice at a frequency of 1.9 GHz. The chamber was characterized for RF dose rate as a function of animal body mass and dose rate variations due to animal movement in the cage. Dose rates were quantified in terms of whole‐body average (WBA) specific absorption rate (SAR), brain average (BA) SAR and peak spatial‐average (PSA) SAR using measurement and computational methods. Measurements were conducted on mouse cadavers in a multitude of possible postures and positions to evaluate the variations of WBA‐SAR and its upper and lower bounds, while computations utilizing the finite‐difference time‐domain method together with a heterogeneous mouse model were performed to determine variations in BA‐SAR and the ratio of PSA‐SAR to WBA‐SAR. Measured WBA‐SAR variations were found to be within the ranges of 9–23.5 W/kg and 5.2–13.8 W/kg per 1 W incident power for 20 and 40 g mice, respectively. Computed BA‐SAR variations were within the ranges of 3.2–10.1 W/kg and 3.3–9.2 W/kg per 1 W incident power for 25 and 30 g mouse models, respectively. Ratios of PSA‐SAR to WBA‐SAR, averaged over 0.5 mg and 5 mg tissue volumes, were observed to be within the ranges of 6–15 and 4–10, respectively. Bioelectromagnetics 33:575–584, 2012. © 2012 Wiley Periodicals, Inc.     

Design and Dosimetric Characterization of a Broadband Exposure Facility for In Vitro Experiments in the Frequency Range 18–40.5 GHz

A novel exposure facility for exposing cell monolayers to centimeter and millimeter waves (18–40.5 GHz) used by future 5G mobile communication technology and similar applications has been developed. A detailed dosimetric characterization of the apparatus for frequencies of 27 and 40.5 GHz and 60 mm petri dishes, used in a presently ongoing study on human dermal fibroblasts and keratinocytes, was carried out. The exposure facility enables a well-defined, randomized, and blinded application of sham exposure and exposure with selectable values of incident power flux density, and additionally provides the possibility of continuous monitoring of the sample temperature during exposure while it does not require significant deviations from routine in vitro handling procedures, i.e. petri dishes are not required to be placed inside waveguides or TEM cells. Mean specific absorption rate (SAR) values inside the cell monolayer of 115 W/kg (27 GHz) and 160 W/kg (40.5 GHz) per watt antenna input power and corresponding transmitted power density (S_t) values at the bottom of the cell monolayer of 65 W/m² (27 GHz) and 70 W/m² (40.5 GHz) per watt antenna input power can be achieved, respectively. For reasonable amounts of harvested cells (80% of petri dish bottom area), the variation (max/min) of SAR and S_t over the cell monolayer remains below 3.7 dB (27 GHz) and 3.0 dB (40.5 GHz), respectively. © 2021 Bioelectromagnetics Society.     

Effects of continuous and intermittent exposure to RF fields with a wide range of SARs on cell growth, survival, and cell cycle distribution

To examine the biological effects of radio frequency (RF) electromagnetic fields in vitro, we have examined the fundamental cellular responses, such as cell growth, survival, and cell cycle distribution, following exposure to a wide range of specific absorption rates (SAR). Furthermore, we compared the effects of continuous and intermittent exposure at high SARs. An RF electromagnetic field exposure unit operating at a frequency of 2.45 GHz was used to expose cells to SARs from 0.05 to 1500 W/kg. When cells were exposed to a continuous RF field at SARs from 0.05 to 100 W/kg for 2 h, cellular growth rate, survival, and cell cycle distribution were not affected. At 200 W/kg, the cell growth rate was suppressed and cell survival decreased. When the cells were exposed to an intermittent RF field at 300 W/kg(pk), 900 W/kg(pk) and 1500 W/kg(pk) (100 W/kg(mean)), no significant differences were observed between these conditions and intermittent wave exposure at 100 W/kg. When cells were exposed to a SAR of 50 W/kg for 2 h, the temperature of the medium around cells rose to 39.1 degrees C, 100 W/kg exposure increased the temperature to 41.0 degrees C, and 200 W/kg exposure increased the temperature to 44.1 degrees C. Exposure to RF radiation results in heating of the medium, and the thermal effect depends on the mean SAR. Hence, these results suggest that the proliferation disorder is caused by the thermal effect.     

A new in vitro exposure device for the mobile frequency of 900 MHz

A wire patch cell has been designed for exposing cell cultures during in vitro experiments studying possible effects of mobile radio telephone. It is based on the wire patch antenna which works at 900 MHz with a highly homogeneous field inside the antenna cavity. The designed cell structure is symmetric and provides a rather homogeneous field distribution in a large area around its centre. Moreover, the exposure cell can irradiate equally up to eight 35 mm Petri dishes at the same time, which enhances the statistical biological studies. To improve the specific absorption rate (SAR) homogeneity inside each sample, each dish is placed into another 50 mm dish. This way, SAR inhomogeneity is always proper for biological studies (below 30%). The main advantage of this new device is that it can provide SAR levels 20 times higher than those induced by classical Crawford transverse electromagnetic (TEM) cell. Moreover, this small open device is easy to construct and fits into an incubator. However, to be used for in vitro, the wire patch cell is a radiating element with the same radiating pattern as a dipole, and thus some absorbing materials are necessary around the system when used for in vitro experiments. Secondly, because of its narrow bandwidth, it is difficult to maintain its working frequency. To overcome this problem, a matching device is integrated into the test cell. In this paper, we present a detailed explanation of the cell behavior and dosimetric assessments for eight 35 mm Petri dishes exposed. Simulations using the Finite Difference Time Domain technique and experimental investigations have been carried out to design the cell at 900 MHz. The numerical dosimetry was validated by dosimetric measurements. These investigations estimated the dosimetric precision at 11%.     

Radiofrequency radiation at 2.856 GHz does not affect key cellular endpoints in neuron-like PC12 cells

To investigate the potential cytotoxicity of radiofrequency (RF) radiation on central nervous system, rat pheochromocytoma (PC12) cells were exposed to 2.856 GHz RF radiation at a specific absorption rate (SAR) of 4 W/kg for 8 h a day for 2 days in 35 mm Petri dishes. During exposure, the real-time variation of the culture medium temperature was monitored in the first hour. Reactive oxygen species (ROS) production, intracellular Ca²⁺ concentration, and cell apoptosis rate were assessed immediately after exposure by flow cytometry. The results showed that the medium temperature raised about 0.93 °C, but no significant changes were observed in apoptosis, ROS levels or intracellular Ca²⁺ concentration after treatment. Although several studies suggested that RF radiation does indeed cause neurological effects, this study presented inconsistent results, indicating that 2.856 GHz RF radiation exposure at a SAR of 4 W/kg does not have a dramatic impact on PC12 cells, and suggests the need for further investigation on the key cellular endpoints of other nerve cells after exposure to RF radiation.     

Low-level microwave irradiation and central cholinergic activity: a dose-response study

Rats were irradiated with circularly polarized, 2,450-MHz pulsed microwaves (2-microseconds pulses, 500 pulses per second [pps]) for 45 min in the cylindrical waveguide system of Guy et al:(Radio Sci 14:63-74, 1979). Immediately after exposure, sodium-dependent high-affinity choline uptake, an indicator of cholinergic activity in neural tissue, was measured in the striatum, frontal cortex, hippocampus, and hypothalamus. The power density was set to give average whole-body specific absorption rates (SAR) of 0.3, 0.45, 0.6, 0.75, 0.9, or 1.2 W/kg to study the dose-response relationship between the rate of microwave energy absorption and cholinergic activity in the different areas of the brain. Decrease in choline uptake was observed in the striatum at a SAR of 0.75 W/kg and above, whereas for the frontal cortex and hippocampus, decreases in choline uptake were observed at a SAR of 0.45 W/kg and above. No significant effect was observed in the hypothalamus at the irradiation power densities studied. The probit analysis was used to determine the SAR50 in each brain area, i.e., the SAR at which 50% of maximum response was elicited. SAR50 values for the striatum, frontal cortex, and hippocampus were 0.65, 0.38, and 0.44 W/kg, respectively.     

Effects of modulated microwave radiation at cellular telephone frequency (1.95 GHz) on X-ray-induced chromosome aberrations in human lymphocytes in vitro

The case for a DNA-damaging action produced by radiofrequency (RF) signals remains controversial despite extensive research. With the advent of the Universal Mobile Telecommunication System (UMTS) the number of RF-radiation-exposed individuals is likely to escalate. Since the epigenetic effects of RF radiation are poorly understood and since the potential modifications of repair efficiency after exposure to known cytotoxic agents such as ionizing radiation have been investigated infrequently thus far, we studied the influence of UMTS exposure on the yield of chromosome aberrations induced by X rays. Human peripheral blood lymphocytes were exposed in vitro to a UMTS signal (frequency carrier of 1.95 GHz) for 24 h at 0.5 and 2.0 W/kg specific absorption rate (SAR) using a previously characterized waveguide system. The frequency of chromosome aberrations was measured on metaphase spreads from cells given 4 Gy of X rays immediately before RF radiation or sham exposures by fluorescence in situ hybridization. Unirradiated controls were RF-radiation- or sham-exposed. No significant variations due to the UMTS exposure were found in the fraction of aberrant cells. However, the frequency of exchanges per cell was affected by the SAR, showing a small but statistically significant increase of 0.11 exchange per cell compared to 0 W/kg SAR. We conclude that, although the 1.95 GHz signal (UMTS modulated) does not exacerbate the yield of aberrant cells caused by ionizing radiation, the overall burden of X-ray-induced chromosomal damage per cell in first-mitosis lymphocytes may be enhanced at 2.0 W/kg SAR. Hence the SAR may either influence the repair of X-ray-induced DNA breaks or alter the cell death pathways of the damage response.     

Comparison of dose dependences for bioeffects of continuous-wave and high-peak power microwave emissions using gel-suspended cell cultures

The study compared bioeffects of continuous wave (CW) microwaves and short, extremely high power pulses (EHPP) at the same carrier frequency (9.3 GHz) and average power (1.25 W). The peak transmitted power for EHPP was 250 kW (0.5-micro s pulse width, 10 p.p.s.), producing the E field of 1.57 MV/m in the waveguide. A biological endpoint was the density of yeast cells, achieved after a 6 h growth period in a solid nutrient medium (agarose gel) during EHPP or CW exposure. Owing to power losses in the medium, the specific absorption rate (SAR) ranged from 3.2 kW/kg at the exposed surface of the sample to 0.6 mW/kg at 24 mm depth. Absorption and penetration of EHPP was identical to CW, producing peak SAR values 200 000 times higher than the average SAR, as high as 650 MW/kg at the surface. CW and EHPP exposures produced highly nonuniform but identical heating patterns in exposed samples. Following the exposure, the samples were sliced in a plane perpendicular to the wave propagation, in order to separate cell masses exposed at different SAR levels. Cell density in the slices was determined by nephelometry and compared to unexposed parallel control samples. Cell density was strongly affected by irradiation, and the changes correlated well with the local temperature rise. However, the data revealed no statistically significant difference between CW and EHPP samples across the entire studied range of SAR levels (over six orders of magnitude). A trend (P<0.1) for such a difference was observed in slices that were exposed at a time average SAR of 100 W/kg and higher, which corresponded to peak SAR above 20 MW/kg for the EHPP condition. These numbers could be indicative of a threshold for a specific (not merely thermal) exposure effect if the trend is confirmed by future studies.     

Ornithine decarboxylase activity of L929 cells after exposure to continuous wave or 50 Hz modulated radiofrequency radiation--a replication study

A replication study with some extensions was made to confirm enhancement of ornithine decarboxylase (ODC) activity in murine L929 fibroblasts after radiofrequency (RF) field exposure reported in earlier studies. L929 cells purchased from two cell banks were exposed for 2, 8, or 24 h to continuous wave or DAMPS (burst modulated at 50 Hz, with 33% duty cycle) signals at specific absorption rate (SAR) levels of 2.5 or 6.0 W/kg. Exposures were carried out in Crawford and waveguide chambers, at frequencies 835 and 872 MHz, respectively. The results did not confirm findings of previous studies reporting increased ODC activity in RF-exposed cells. When Crawford cell exposure system was used, ODC activity was either not affected (in the case of 8 or 24 h exposures) or decreased after 2 h exposure at the highest SAR level (6 W/kg). The decrease was most pronounced when cooling with air flow was not used, and is most likely related to increased temperature. The minor methodological differences (use of antibiotics, increased sensitivity of ODC assay) are not likely to explain the inconsistency of the findings of the present and previous studies. Different results were obtained in experiments with the waveguide system that involves more efficient temperature control. In this exposure system, ODC activity was increased after 8 h exposure at 6 W/kg. Further studies are warranted to explore whether this finding reflects a true non-thermal effect. The present study did not provide evidence for modulation-specific effects reported in earlier studies.     

Effects of high frequency electromagnetic fields on micronucleus formation in CHO-K1 cells

To investigate the effects of high frequency electromagnetic fields (HFEMFs), we assessed the frequency of micronucleus (MN) formation induced by chromosomal breakage or inhibition of spindles during cell division in Chinese hamster ovary (CHO)-K1 cells, using the cytokinesis block micronucleus method. The MN frequency in cells in the inner, middle and outer wells of an annular culture plate was determined for the following four conditions: (1) CHO-K1 cells were exposed to a HFEMF for 18 h at average specific absorption rates (SARs) of 13, 39 and 50 W/kg with input power 7.8 W, and were compared with a sham-exposed control; (2) the cells were also exposed to a HFEMF at SARs of 78 and 100 W/kg with input power 13 W, and were compared with a sham-exposed control; (3) the cells were treated with bleomycin alone or with bleomycin followed by exposure to a HFEMF for 18 h at SARs of 25, 78 and 100 W/kg, and were compared with a bleomycin-treated positive control. The cells treated with bleomycin alone were compared with sham-exposed controls; and (4) As a high temperature control, CHO-K1 cells were incubated at 39 degrees C for 18 h. In study (1), the MN frequency of cells exposed to a HFEMF at a SAR of up to 50 W/kg was not different to that in sham-exposed cells. In study (2), there were statistically significant increases in the MN frequencies of cells in the middle and outer wells of the annular culture plate caused by exposure to a HFEMF at 100 and 78 W/kg, respectively. In study (3), the MN frequencies of cells in the middle (100 W/kg) and outer wells (78 W/kg) of the annular culture plate were statistically higher than that caused by bleomycin-treatment alone. In study (4), there was a statistically significant increase of MN frequency in the cells treated by heat at 39 degrees C.These results indicate that cells exposed to a HFEMF at a SAR of 78 W/kg and higher form MN more frequently than sham-exposed cells, while exposure to a HFEMF at up to 50 W/kg does not induce MN formation. In addition, a HFEMF at a SAR of 78 W/kg and higher may potentiate MN formation induced by bleomycin-treatment.     

Implantation in vitro: co-culture of rat blastocyst and epithelial cell vesicles

Implantation of blastocysts involves conversion of maternal and embryonic cell surfaces from a nonadhesive to an adhesive state in response to the internally driven developmental program or to externally generated factors. However, the intricacies of the cellular and subcellular changes that promote the attachment are not known, because these changes are difficult to determine in situ because of the nonaccessibility of the site. To overcome this, an in vitro model of implantation was developed by co-culturing rat blastocysts and uterine epithelial cells of the same gestational age (day 5 postcoitum; plug day as day 1) in drops hanging from the lid of a Petri dish. The system was used to study the changes on the surface membranes of the cells of the trophectoderm and uterine epithelium and to evaluate the antiadhesive activity of the newly designed test substances. The isolated epithelial cell vesicles were co-cultured with zona-free blastocysts in the microdrops (40–50 µl) hanging from the lid of a 60-mm Petri dish. The lid was placed over the lower dish, which was presaturated with the medium. The culture was examined 48 h later to determine the site of adhesion of epithelial cell vesicles with the trophoblasts lining the blastocyst. The cell-cell adhesion was monitored on a computerized image analyzer. To validate the adhesion of blastocysts and epithelial cell vesicles in co-culture, the expression of a cell adhesion molecule, uvomorulin, was studied using immunocytochemical technique after incubating with antiuvomorulin antibody. Intense staining was noted on the membrane surfaces at the site of attachment of the blastocyst and cell vesicles.The authors express their sincere thanks to the Ministry of Health and Family Welfare, Government of India, for their financial support     

Temperature regulation in the unrestrained rabbit during exposure to 600 MHz radiofrequency radiation

Six male New Zealand white rabbits were individually exposed to 600 MHz radiofrequency (RF) radiation for 90 min in a waveguide exposure system at an ambient temperature (Ta) of 20 or 30 degrees C. Immediately after exposure, the rabbit was removed from the exposure chamber and its colonic and ear skin temperatures were quickly measured. The whole-body specific absorption rate (SAR) required to increase colonic and ear skin temperature was determined. At a Ta of 20 degrees C the threshold SAR for elevating colonic and ear skin temperature was 0.64 and 0.26 W/kg, respectively. At a Ta of 30 degrees C the threshold SARs were slightly less than at 20 degrees C, with values of 0.26 W/kg for elevating colonic temperature and 0.19 W/kg for elevating ear skin temperature. The relationship between heat load and elevation in deep body temperature shown in this study at 600 MHz is similar to past studies which employed much higher frequencies of RF radiation (2450-2884 MHz). On the other hand, comparison of these data with studies on exercise-induced heat production and thermoregulation in the rabbit suggest that the relationship between heat gain and elevation in body temperature in exercise and from exposure to RF radiation may differ considerably. When combined with other studies, it was shown that the logarithm of the SAR required for a 1.0 degree C elevation in deep body temperature of the rabbit, rat, hamster, and mouse was inversely related to the logarithm of body mass. The results of this study are consistent with the conclusion that body mass strongly influences thermoregulatory sensitivity of the aforementioned laboratory mammals during exposure to RF radiation.     

In utero exposure to microwave radiation and rat brain development

Timed-pregnancy rats were exposed in a circular waveguide system starting on day 2 of gestation. The system operated at 2,450 MHz (pulsed waves; 8 microseconds PW; 830 pps). Specific absorption rate (SAR) was maintained at 0.4 W/kg by increasing the input power as the animals grew in size. On day 18 of gestation the dams were removed from the waveguide cages and euthanized; the fetuses were removed and weighed. Fetal brains were excised and weighed, and brain RNA, DNA and protein were determined. Values for measured parameters of the radiated fetuses did not differ significantly from those of sham-exposed fetuses. A regression of brain weight on body weight showed no micrencephalous fetuses in the radiation group when using as a criterion a regression line based on two standard errors of the estimate of the sham-exposed group. In addition, metrics derived from brain DNA (ie, cell number and cell size) showed no significant differences when radiation was compared to sham exposure. We conclude that 2,450-MHz microwave radiation, at an SAR of 0.4 W/kg, did not produce significant alterations in brain organogenesis.     

Reevaluation and improved design of the TEM cell in vitro exposure unit for replication studies

The transverse electromagnetic (TEM) cell system developed by Litovitz et al. and utilized by Penafiel et al. for the exposure of cells in T25 flasks at 835 MHz has been reevaluated for the purpose of replicating the studies published by Penafiel. The original setup has been reconstructed as closely as possible, with improvements enabling blinded exposures, forced cooling and better repeatable positioning of the flasks, as well as tight exposure and environmental parameter control. The signal unit can simulate the original signal but also enables various other exposure schemes. The setup has been evaluated for four T25 flasks filled with 5 and 10 ml of cell medium by experimental and numerical means. Comparing E field, SAR and temperature measurements resulted in good agreement: <0.4 dB (4.5%) for E field and 0.48 dB (10.5%) for SAR. The overall average SAR within the medium is 6.0 W/kg at 1 W input power with a standard deviation of less than 52%. The temperature increase was determined to be 0.13 degrees C/(W/kg). This can be reduced to 0.045 degrees C/(W/kg) by applying active air flow cooling. The comparison of SAR values from temperature measurements with the corresponding simulated values resulted in excellent agreement. These results do not correspond to the previous study reporting an average SAR within the medium of 2.5 W/kg at an input power of 0.96 W.     

A simple method for reducing moisture condensation on Petri dish lids

Condensation on the lids of Petri dishes, used to culture plant tissues, can often obscure the view of the contents of the dish and interfere with data collection. Under the high humidity conditions that exist in the culture container, a small temperature drop causes water to condense on the inside lid and sides of the container. Mild condensation causes “fogging” while continual or repeated rounds of condensation result in the formation of water droplets. To control condensation in the standard plant tissue culture Petri dish, a simple method was developed whereby the lid of the culture dish was modified, to buffer the lid from temperature fluctuations. Polymer discs, which were the same diameter as the Petri dish lid, were either placed on the top of the lids of existing dishes or surface-sterilized and used in place of the lid. Polymer discs of varying thicknesses and type, and possessing different thermal conductivities, were evaluated for their abilities to reduce the rate of condensation formation. Petri dishes with modified lids were placed under reduced temperature conditions. Condensation, forming on the lids of the dishes was quantified over time using image analysis. Gray value determinations indicated that the thicker polymer discs with the lowest thermal conductivities provided the best protection against condensation. Placement of polymer discs on the top of Petri dishes is a relatively simple method that can be used to buffer the lid from small temperature changes and minimize condensation problems.