SAR versus VAR, and the size and shape that provide the most appropriate RF exposure metric in the range of 0.5-6 GHz

Basic restrictions for protecting against localized tissue heating induced from exposure to radiofrequency (RF) fields are typically specified as the specific energy absorption rate (SAR), which is mass averaged in recognition of the thermal diffusion properties of tissues. This article seeks to determine the most appropriate averaging mass (1, 3, 5, 7, or 10 g) and averaging shape (cube or sphere). We also consider an alternative metric, volumetric energy absorption rate (VAR), which uses volume averaging (over 1, 3, 5, 7, and 10 cm³; cube and sphere). The SAR and VAR averaging approaches were compared by considering which was a better predictor of tissue temperature rise (ΔT) induced by near‐ and far‐field RF exposures (0.5–6 GHz), calculated in a detailed human body model. For the exposure scenarios that we examined, VAR is better correlated with ΔT than SAR, though not at a statistically significant level for most of the metric types we studied. However, as VAR offers substantive advantages in ease of assessment we recommend this metric over SAR. Averaging over a cube or a sphere provides equivalent levels of correlation with ΔT, and so we recommend choosing the averaging shape on the basis of which is easier to assess. The optimal averaging volume is 10 cm³ for VAR, and the optimal mass is 10 g for SAR. The correlation between VAR or SAR and ΔT diminishes substantially at 6 GHz, where incident power flux density may be a better exposure metric. Bioelectromagnetics 32:312–321, 2011. © 2010 Wiley‐Liss, Inc.     

Near-field dosimetry for in vitro exposure of human cells at 60 GHz

Due to the expected mass deployment of millimeter‐wave wireless technologies, thresholds of potential millimeter‐wave‐induced biological and health effects should be carefully assessed. The main purpose of this study is to propose, optimize, and characterize a near‐field exposure configuration allowing illumination of cells in vitro at 60 GHz with power densities up to several tens of mW/cm². Positioning of a tissue culture plate containing cells has been optimized in the near‐field of a standard horn antenna operating at 60 GHz. The optimal position corresponds to the maximal mean‐to‐peak specific absorption rate (SAR) ratio over the cell monolayer, allowing the achievement of power densities up to 50 mW/cm² at least. Three complementary parameters have been determined and analyzed for the exposed cells, namely the power density, SAR, and temperature dynamics. The incident power density and SAR have been computed using the finite‐difference time‐domain (FDTD) method. The temperature dynamics at different locations inside the culture medium are measured and analyzed for various power densities. Local SAR, determined based on the initial rate of temperature rise, is in a good agreement with the computed SAR (maximal difference of 5%). For the optimized exposure setup configuration, 73% of cells are located within the ±3 dB region with respect to the average SAR. It is shown that under the considered exposure conditions, the maximal power density, local SAR, and temperature increments equal 57 mW/cm², 1.4 kW/kg, and 6 °C, respectively, for the radiated power of 425 mW. Bioelectromagnetics 33:55–64, 2012. © 2011 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.     

Operant behavior and colonic temperature of Macaca mulatta exposed to radio frequency fields at and above resonant frequencies

Five food-deprived rhesus monkeys were exposed to 225-MHz continuous-wave, and 1.3-GHz, and 5.8-GHz pulsed radiation to determine the minimal power densities affecting performance. The monkeys were trained to press a lever (observing-response) thereby producing signals that indicated availability of food. In the presence of the aperiodically appearing food signals, a detection response on a different lever was reinforced by a food pellet. Continuous, stable responding during 60-min sessions developed and was followed by repeated exposures to radiofrequency radiation. The subjects, restrained in a Styrofoam chair, were exposed to free-field radiation while performing the task. Colonic temperature was simultaneously obtained. Observing-response performance was impaired at increasingly higher power densities as frequency increased from the near-resonance 225 MHz to the above-resonance 5.8 GHz. The threshold power density of disrupted response rate at 225 MHz was 8.1 mW/cm²; at 1.3 GHz it was 57 mW/cm², and at 5.8 GHz it was 140 mW/cm². These power densities were associated with reliable increases in colonic temperatures above sham-exposure levels. The mean increase was typically in the range of 1°C, and response-rate changes were not observed in the absence of concomitant temperature increases. In these experiments increase of colonic temperature was a much better predictor of behavioral disruption than was either the power density of the incident field or estimates of whole-body-averaged rates of energy absorption.     

Current density in a model of a human body with a conductive implant exposed to ELF electric and magnetic fields

A numerical model of a human body with an intramedullary nail in the femur was built to evaluate the effects of the implant on the current density distribution in extremely low frequency electric and magnetic fields. The intramedullary nail was chosen because it is one of the longest high conductive implants used in the human body. As such it is expected to alter the electric and magnetic fields significantly. The exposure was a simultaneous combination of inferior to superior electric field and posterior to anterior magnetic field both alternating at 50 Hz with the values corresponding to the ICNIRP reference levels: 5000 V m^?1 for electric field and 100 µT for magnetic flux density. The calculated current density distribution inside the model was compared to the ICNIRP basic restrictions for general public (2 mA m^?2). The results show that the implant significantly increases the current density up to 9.5 mA m^?2 in the region where it is in contact with soft tissue in the model with the implant in comparison to 0.9 mA m^?2 in the model without the implant. As demonstrated the ICNIRP basic restrictions are exceeded in a limited volume of the tissue in spite of the compliance with the ICNIRP reference levels for general public, meaning that the existing safety limits do not necessarily protect implanted persons to the same extent as they protect people without implants. Bioelectromagnetics 30:591–599, 2009. © 2009 Wiley‐Liss, Inc.     

Microwaves modify thermoregulatory behavior in squirrel monkey

Squirrel monkeys (Saimiri sciureus) trained to regulate environmental temperature (T_a) behaviorally were exposed in the far field of a horn antenna to ten-minute periods of 2,450 MH_z CW microwaves. Incident power density ranged from 1 to 22 mW/cm². The corresponding specific absorption rate (SAR), derived from temperature increments in saline-filled styrofoam models, ranged from 0.15 to 3.25 W/kg. Controls included exposure to infrared radiation of equivalent incident energy and no radiation exposure. Normal thermoregulatory behavior produces tight control over environmental and body temperatures; most monkeys select a T_a of 34–36°C. Ten-minute exposures to 2,450 MH_z CW microwaves at an incident power density of 6–8 mW/cm² stimulated all animals to select a lower T_a. This threshold energy represents a whole-body SAR of 1.1 W/kg, about 20% of the resting metabolic rate of the monkey. Thermoregulatory behavior was highly efficient, and skin and rectal temperatures remained stable, even at 22 mW/cm² where the preferred T_a was lowered by as much as 4°C. No comparable reduction in selected T_a below control levels occurred during exposure to infrared radiation of equal incident power density.     

Evaluation of the Potential In Vitro Antiproliferative Effects of Millimeter Waves at Some Therapeutic Frequencies on RPMI 7932 Human Skin Malignant Melanoma Cells

The potential antiproliferative effects of low power millimeter waves (MMWs) at 42.20 and 53.57 GHz on RPMI 7932 human skin melanoma cells were evaluated in vitro in order to ascertain if these two frequencies, comprised in the range of frequency used in millimeter wave therapy, would have a similar effect when applied in vivo to malignant melanoma tumours. Cells were exposed for 1 h exposure/day and to repeated exposure up to a total of four treatments. Plane wave incident power densities <1 mW/cm² were used in the MMWs-exposure experiments so that the radiations did not cause significant thermal effects. Numerical simulations of Petri dish reflectivity were made using the equations for the reflection coefficient of a multilayered system. Such analysis showed that the power densities transmitted into the aqueous samples were ≤0.3 mW/cm². Two very important and general biological endpoints were evaluated in order to study the response of melanoma cells to these radiations, i.e. cell proliferation and cell cycle. Herein, we show that neither cell doubling time nor the cell cycle of RPMI 7932 cells was affected by the frequency of the GHz radiation and duration of the exposure, in the conditions above reported.     

Effects of Microwave Exposure at Various Power Densities on Mitochondrial Marker Enzymes in Mouse Brains

Four groups of C₅₇BL mice were irradiated with 3 GHz pulse (PW) microwaves for 3 hours at incident power densities of 0.1, 0.5, 1 and 5 mW/cm² respectively. The amount of mitochondria1 marker enzymes succinate dehydrogenase (SDH) and monoamine oxidase (MAO) in the hypothalamus and hippocampus were determined by microspectrophotometry. SDH and MA0 in the irradiated groups (except 0.1 mW/cm²) were significantly lower compared to the control group (p < 0.01). The lowest level occurred in the 5 mW/cm² group. The threshold level was 0.5 mW/cm². To compare the effects of PW with continuous wave (CW) exposure, two experimental groups were exosed to 2.45 GHz, using CW; the enzymes were decreased only in the 5 mW/cm² group. The results show that PW radiation is more effective then CW radiation in decreasing SDH and MA0 levels.     

Resonance effect of millimeter waves in the power range from 10-19 to 3 × 10-3 W/cm2 on Escherichia coli cells at different concentrations

The effect of millimeter waves (MMWs) on the genome conformational state (GCS) of E. coli AB1157 cells was studied by the method of anomalous viscosity time dependencies (AVTD) in the frequency range of 51.64-51.85 GHz. The 51.755 GHz resonance frequency of the cell reaction to MMWs did not depend on power density (PD) in the range from 10^-19 to 3 × 10^-3 W/cm². The half-width of the resonant reaction of cells showed a sigmoid dependence on PD, changing from 3 MHz to 100 MHz. The PD dependence of the half-width had the same shape for different concentrations of exposed cells (4 × 10⁷ and 4 × 10⁸ cells/ml), whereas the magnitude of the 51.755 GHz resonance effect differed significantly and depended on the PD of MMW exposure. Sharp narrowing of the 51.755 GHz resonance in the PD range from 10^-4 to 10^-7 W/cm² was followed by an emergence of new resonance frequencies. The PD dependence of the MMW effect at one of these resonance frequencies (51.674 GHz) differed markedly from the corresponding dependence at the 51.755 GHz resonance, the power window occurring in the range from 10^-16 to 10^-8 W/cm². The results obtained were explained in the framework of a model of electron-conformational interactions. The frequency-time parameters of this model appeared to be in good agreement with experimental data. © 1996 Wiley-Liss, Inc.     

Cooperative Response of Escherichia Coli Cells to the Resonance Effect of Millimeter Waves at Super Low Intensity

Cells of Escherichia coli K12 AB1157 were irradiated with millimeter waves (MMW) within the power density (PD) range of 10^?20 to 10 ⁴ W/cm². MMW were applied for 0.5–70 min at 51.76 GHz or 41.32 GHz at which, as had been shown earlier, MMW resonantly changes the genome conformational state (GCS) of E. coli K12 AB1157 cells. The changes in the GCS were tested with the method of anomalous viscosity time dependence (AVTD). It was demonstrated that the resonance effect of MMW manifests itself at PD up to 10^?19 W/cm². Dependences of MMW effect on power density and time of exposure proved to have distinct characteristics when cells are irradiated during the logarithmic or stationary phases of the culture's growth. It was found that the resonance effect of MMW on the GCS of E. coli cells at the early stationary phase changes the developmental dynamics of the irradiated culture. It was established for the first time that the magnitude of the resonance MMW effect depends on the concentration of irradiated cells. An analysis of the results indicates an electromagnetic rather than diffusion nature of the cells' cooperative responses to millimeter waves.     

Modification of the repeated acquisition of response sequences in rats by low-level microwave exposure

The acute effects of microwave exposure on a repeated acquisition baseline were investigated in three rats. Each session the animals acquired a different four-member response sequence. Each of the first three correct responses advanced the sequence to the next member, and the fourth correct response produced food reinforcement. Incorrect responses produced a three-second timeout. Baseline and control sessions were characterized by a decrease in errors within each session. The animals were acutely exposed to a 2.8 GHz pulsed-microwave field prior to test sessions, with average power densities ranging from 0.25 to 10 mW/cm². In comparison to control sessions, 1/2 hour of exposure to microwave radiation at power densities of 5 and 10 mW/cm² increased errors and altered the pattern of within-session acquisition. Exposure to the 10 mW/cm² power density decreased the rate of sequence completion in all animals. The results of exposures at 0.25, 0.5, and 1 mW/cm² power densities were generally within the control range. The results are interpreted as indicating a disruption in the discriminative stimulus control of the repeated acquisition behavior.     

Chronic exposure of rabbits to 0.5 and 5 mW/cm2 2450-MHz CW Microwave Radiation

Two groups of 16 male New Zealand rabbits were exposed to 2450-MHz continuous wave microwave fields in two experiments of 90 days each. The incident power densities of the first and second experiment were 0.5 and 5 mW/cm², respectively. During each study, 16 animals were adapted to a miniature anechoic chamber exposure system for at least 2 weeks, then 8 of them were exposed for 7 h daily, 5 days a week for 13 weeks, and the other 8 animals were sham exposed. The rabbits were placed in acrylic cages, and each was exposed from the top in an individual miniature anechoic chamber. Thermography showed a maximum specific absorption rate of 5.5 W/kg in the head and 7 W/kg in the back at 5-mW/cm² incident power density. After each 7-h exposure session, the animals were returned to their home cages. Food consumption in the exposure chamber and body mass were measured daily. Blood samples were taken before exposure and monthly thereafter for hematological, morphological, chemical, protein electrophoresis, and lymphocyte blast transformation studies. Eyes were examined for cataract formation. Finally, pathological examinations of 28 specimens of organs and tissues of each rabbit were performed. Statistically, there was a significant (P < .01) decrease only of food consumption during the 5-mW/cm² exposure; other variables were not significantly different between exposed and control groups.     

Sialolithiasis: Application parameters for an optimal laser therapy

In‐vitro experimental parametric studies of laser ablation using natural sialoliths and artificial stones have been performed toward an efficient laser treatment of sialolithiasis. Surface microstructure and water adsorption become critical for coupling high power pulsed Ho:YAG laser radiation (λ = 2080 nm, τ ～250 μsec), inducing ablative interactions and stone fragmentation. Results reveal a generic trend, with single pulse laser energy density threshold for sialolith ablative erosion at ～200 J cm^?2 (corresponding to intensity ～800 kW cm^?2) and fragmentation rates reaching ～1 mm/pulse at ～2400 J cm^?2. This process shows no saturation, suggesting that very high energy density irradiation at low pulse repetition rate is an efficient approach. Such operation facilitates rapid cooling and minimal thermal loading of the oral and maxillofacial area, thus causing negligible adverse effects. The method is expected to contribute to the establishment of an easy and optimal therapeutic protocol for sialolithiasis pathology.     

The in vivo effects of 2.45 GHz microwave radiation on rabbit serum components and sleeping times

Summary An investigation was conducted to determine the effects of relatively low power density microwave exposures on various serum components of the Dutch rabbit. Both continuous wave and pulsed mode exposures at 2.45 GHz were used at power densities of 25, 10 and 5 mW/cm². Studies of 10 serum components were performed. Additional studies were conducted on changes in sleeping times of pentobarbital-sedated rabbits at various power densities. Gross and histopathological examinations were performed on representative samples of animals.Changes in the blood chemistry of irradiated animals were consistent with a dose-dependent response to a non-specific thermal stress at all power densities used. Observed physiological response, as well as rectal temperature measurements, indicated that the thermoregulatory capability of the rabbits was sufficient to compensate for the thermal burden at 5 and 10 mW/cm², but could be overridden by a 2 h exposure at 25 mW/cm². Pathology findings included a mild, repairable nephrosis in animals exposed at a power density of 25 mW/cm².A further investigation of analeptic effects at power densities varying from 5 mW/cm² to 50 mW/cm² resulted in a statistically significant decrease in sleeping times, apparently proportional to power density below 15 mW/cm².This research was partially supported by the US Army Medical Research and Development Command, Contract No. DADA17-72-C-2144. (The views expressed are those of the authors and do not necessarily reflect those of the Department of the Army)     

Observing-responses of rats exposed to 1.28- and 5.62-GHz microwaves

The effects of microwave irradiation at two different frequencies (1.28 and 5.62 GHz) on observing-behavior of rodents were investigated. During daily irradiation, eight male hooded rats performed on a two-lever task; depression of one lever produced one of two different tones and the other lever produced food when depressed in the presence of the appropriate tone. At 5.62 GHz, the observing-response rate was not consistently affected until the power density approximated 26 mW/cm² at 1.28 GHz, the observing-response rate of all rats was consistently affected at a power density of 15 mW/cm². The respective whole-body specific absorption rates (SARs) were 4.94 and 3.75 W/Kg. Measurements of localized SAR in a rat-shaped model of simulated muscle tissue revealed marked differences in the absorption pattern between the two frequencies. The localized SAR in the model's head at 1.28 GHz was higher on the side distal to the source of radiation. At 5.62 GHz the localized SAR in the head was higher on the proximal side. It is concluded that the rat's observing behavior is disrupted at a lower power density at 1.28 than at 5.62 GHz because of deeper penetration of energy at the lower frequency, and because of frequency-dependent differences in anatomic distribution of the absorbed microwave energy.     

Real-time measurements of quartz erosion in experiments modeling heat loads on divertor plates during disruption in tokamaks

Time-resolved measurements of quartz erosion are carried out to determine the density of the energy flux incident onto the sample surface. These data are needed to create a reliable code describing the interaction of a thermonuclear plasma with a solid surface. Experiments were performed in the 2MK-200 facilityunder the program of modeling heat loads on divertor plates during disruptions in tokamaks. A 10-mm-thick plate of fused quartz was exposed to a high-temperature deuterium-plasma stream with the temperature T _i +T _e ≤1 keV, density (5–10)×10¹⁵ cm⁻³ β=0.25, energy density up to 200 J/cm², and power density ∼10 MW/cm². It is shown that the quartz erosion begins almost immediately after the stream reaches the surface. The eroded material shields the quartz surface from further destruction. Under the given experimental conditions, the integral shielding factor (the ratio of the stream energy to the energy reaching the surface) was rather high (about seventeen). As a result, at a stream energy density of ∼150 J/cm², the total erosion depth was about 0.75 μm over 35 μs. Based on the measured time dependence of the erosion depth and the reference data on the thermal conductivity of the fused quartz, the power density incident onto the quartz sample was numerically calculated. __________ Translated from Fizika Plazmy, Vol. 27, No. 3, 2001, pp. 243–250. Original Russian Text Copyright ? 2001 by Arkhipov, Bakhtin, Vasenin, Zhitlukhin, Safronov, Toporkov.     

Temperature oscillations in liquid media caused by continuous (nonmodulated) millimeter wavelength electromagnetic irradiation

Convection in liquids caused by 53–78 GHz millimeter wave irradiation with incident power density that ranged from 10 μW/cm² to 1 W/cm² was studied. Infrared thermography was used as an artifact-free method for recording surface-temperature dynamics during irradiation. It was found that continuous (nonmodulated) waves can produce a relaxation-type temperature oscillation in liquids with a relatively high stability of the period between temperature spikes. The temperature oscillation is due to the repetitive formation and dissipation of a torroidal type of convection vortex. When the vortex became stable during irradiation, we observed a temperature decrease following the initial temperature-rise phase, even though the irradiation was constantly maintained. This result constitutes a new process that can play a significant role in producing microwave bioeffects, including some so-called “nonthermal” effects and some effects that are inversely related to heating. Also, it can be considered as a newly discovered potential artifact in microwave bioeffects studies. © 1996 Wiley-Liss, Inc.     

Exposure Assessment in Millimeter-Wave Reverberation Chamber Using Murine Phantoms

This study deals with the design and calibration of the first mode-stirred reverberation chamber (RC) in the 60-GHz-band adapted for in vivo bioelectromagnetic studies. In addition to the interface for electromagnetic and thermal dosimetry, the interfaces for lighting and ventilation were integrated into the RC walls while preserving acceptable shielding. The RC with mechanical and electronic steering capabilities is characterized in the 55–65 GHz range. To this end, murine skin-equivalent phantoms of realistic shape were designed and fabricated. Their complex permittivity is within ±12% of the target value of murine skin (6.19–j5.81 at 60 GHz). The quality factor of the RC loaded with an animal cage, bedding litter, and five murine phantoms was found to be 1.2 × 10⁴. The losses inside the RC were analyzed, and it was demonstrated that the main sources of the power dissipation were the phantoms and mice cage. The input power required to reach the average incident power density of 1 and 5 mW/cm² was found to be 0.23 and 1.14 W, respectively. Surface heating of the mice models was measured in the infrared (IR) range using a specifically designed interface, transparent at IR and opaque at millimeter waves (mmW). Experimental results were compared with an analytical solution of the heat transfer equation and to full-wave computations. Analytical and numerical results were in very good agreement with measurements (the relative deviation after 90 min of exposure was within 4.2%). Finally, a parametric study was performed to assess the impact of the thermophysical parameters on the resulting heating. Bioelectromagnetics. 2020;41:121–135. © 2020 Bioelectromagnetics Society.     

Analysis of medullated axon exposed to radio-frequency electromagnetic radiation

A theoretical model is formulated to determine the electric field and thermal heating produced in a single neuron (medullated axon) by an incident radio-frequency electromagnetic field. The axon is assumed to be embedded in bulk nervous tissue below a planar interface between the tissue and air, with the irradiating field a plane wave normally incident from the air. Heat is removed by blood flow in the tissue. Numerical calculations for incident fields of power density 10mW?cm² and frequencies in the range 10⁸–10¹⁰ Hz show that the oscillating potential difference produced across the cellular membrane (single bilayer) of an unmyelinated axon is less than 5 μV, while that produced across the nodal membrane of a medullated axon may be 2–6 times greater, and that produced across the myelin of a medullated axon about 100 times greater. In the steady state, the temperature differences within the components of the medullated axon are found to be less than 10^-6°C and the temperature gradients less than 0.1°C?cm; these are shown to be negligible.     

Effect of Scalp and Facial Hair on Air Displacement Plethysmography Estimates of Percentage of Body Fat

Objective: The objective of this study was to determine the effect of body hair (scalp and facial) on air displacement plethysmography (BOD POD) estimates of percentage of body fat. Research Methods and Procedures: A total of 25 men (31.4 ± 8.0 years, 83.4 ± 12.2 kg, 181.8 ± 6.9 cm) agreed to grow a beard for 3 weeks to participate in the study. Total body density (g/cm³) and percentage of body fat were evaluated by BOD POD. To observe the effect of trapped isothermal air in body hair, BOD POD measures were performed in four conditions: criterion method (the beard was shaven and a swimcap was worn), facial hair and swimcap, facial hair and no swimcap, and no facial hair and no swimcap_. Results: The presence of only a beard (facial hair and swimcap) resulted in a significant underestimation of percentage of body fat (16.2%, 1.0618 g/cm³) vs. the criterion method (17.1%, 1.0597 g/cm³, p < 0.001). The effect of scalp hair (no swim cap worn) resulted in a significant underestimation in percentage of body fat relative to the criterion method, either with facial hair (facial hair and no swimcap; 14.8%, 1.0649 g/cm³) or without facial hair (no facial hair and no swimcap; 14.8%, 1.0650 g/cm³, p < 0.001 for both). Discussion: A significant underestimation of percentage of body fat was observed with the presence of facial hair (～1%) and scalp hair (～2.3%). This underestimation in percentage of body fat may be caused by the effect of trapped isothermal air in body hair on body‐volume estimates. Thus, excess facial hair should be kept to a minimum and a swimcap should be worn at all times to ensure accurate estimates of body fat when using the BOD POD.