Effect of 5-hydroxytryptamine on tissue blood flow and microwave heating of rat tumors.

We have investigated the effect of 5-hydroxytryptamine (5-HT) on SMT-2A mammary adenocarcinoma blood flow during localized microwave hyperthermia treatment. Tissue blood flow in isogeneic female W/Fu rats was estimated using 25-micron-diameter 113Sn-labeled microspheres. An intraarterial injection of 5-HT (1 mg/kg) into either conscious or anesthetized (Nembutal, 20 mg/kg) animals resulted in a 53% reduction in tumor blood flow, while that of the surrounding skeletal muscle remained unchanged. Because of the selective reduction in tumor perfusion, the blood flow of the normal and malignant tissue was equal after 5-HT injection. This blood flow equivalence remained unaltered after 45 min of heating at 42 degrees C. Consequently, the temperature in the tumor was not significantly different from that in the surrounding normal tissue. In contrast, when the tissues were heated at 42 degrees C without 5-HT, the tumor blood flow was significantly greater than that in the surrounding musculature, resulting in the tumor being 1 degree C lower than the muscle temperature. An intratumoral injection of 5-HT (0.25 mg) reduced the tumor blood flow by 92%, and the blood flow of the surrounding muscle was reduced by 57%. These tissue blood flows were not significantly altered by heating at 44 degrees C for 45 min, and the tumor temperature was 0.7 degrees C greater than that in the muscle. When heating at 44 degrees C was performed without 5-HT injection, the tissue temperatures were equal. Thus, both an intraarterial and an intratumoral injection of 5-HT prior to hyperthermia treatment significantly improved the temperature differential between the neoplastic and surrounding normal tissue. Of additional interest was the observation that an intratumoral injection of 0.15 M NaCl also resulted in a preferential increase in the tumor temperature.

Inhibition of DNA synthesis and enhancement of the uptake and action of methotrexate by low-power-density microwave radiation in L1210 leukemia cells.

We have studied the effects of low-power-density microwave (MW) radiation (continuous and pulsed wave with average power density of 10 milliwatts/sq cm and range of 5 to 50 milliwatts/sq cm; frequency, 1.0 GHz) on the uptake and action of methotrexate (MTX), the inhibition of DNA synthesis in L1210 murine leukemia cells in vitro, and the MTX treatment of mice bearing this leukemia. Using short-term tissue culture techniques, MTX concentrations of 0.2 microM, and MW exposure times of 20 min, we have found that continuous-wave low-power-density MW irradiation enhances the uptake of [3H]MTX as compared to nonirradiated controls. The enhancement is observed in only a small range of power densities (5 to 25 milliwatts/sq cm) and is in an inverted-U-shaped relationship. MW irradiation alone has an inhibitory effect on the [3H]deoxyuridine incorporation into DNA. Compared to cell suspensions treated with MTX alone, groups treated with MW irradiation followed by MTX exhibit an augmentation of inhibition of DNA synthesis as measured by [3H]deoxyuridine incorporation. Combined treatment of L1210-bearing mice with MW irradiation and MTX in vivo prolonged the duration of survival over that of animals treated with MTX alone, indicating a greater killing of leukemia cells. These results suggest that the therapeutic index of MTX may be improved by the use of MW irradiation at low power densities.

Microwave hyperthermia and its effect on tumor blood flow in rats.

The present experiments were designed to investigate the effects of microwave hyperthermia on the microcirculation of normal and malignant tissues in female Wistar/Furth rats. During the localized heating of anesthetized rats, the hind leg musculature surrounding the SMT-2A mammary adenocarcinoma was heated at either 39 degrees, 42 degrees, or 44 degrees. Neither the body temperature, cardiac output, heart rate, nor systemic arterial pressure were significantly altered by heating at these temperatures for up to 60 min. Our results demonstrate that the changes in vascular resistance which occur during hyperthermic treatment are dependent upon both the temperature and the tissue heated. When the tumor (1.2 g)-bearing hind leg was heated to 39 degrees, the tissue vascular resistance and blood flow were unaltered even after 45 min of heating. Heating at 42 degrees and 44 degrees caused an initial vasoconstriction in the tumor, which was subsequently followed by marked vasodilation. This transient initial decrease in blood flow was not, however, observed in the skeletal muscle at either temperature. With prolonged heating at 42 degrees and 44 degrees, the muscle blood flow increased by a factor of 1.6 and 3.2, respectively. In contrast, malignant tissue blood flow increased by a factor of 1.3, and this maximum increase was observed only when the tumor was heated at 44 degrees for more than 45 min. Nevertheless, even with this proportionally greater increase in the blood flow of the surrounding normal tissue, it was never more than that of the tumor. As a consequence, the tumor temperature during hyperthermic treatment was always either less than or equal to that in the surrounding normal musculature.

Immunomagnetic purging of breast cancer from bone marrow for autologous transplantation.

Intensive chemotherapy with autologous bone marrow transplantation is a promising approach for the treatment of breast cancer, provided that clonogenic tumor cells do not contaminate the patient's bone marrow. We have previously demonstrated that a combination of 4-hydroperoxycyclophosphamide (4-HC) and immunomagnetic purging (IMP) with monoclonal antibodies and microspheres could remove 4-5 logs of clonogenic breast cancer cells from a 10-fold excess of human bone marrow cells. In the present report we have evaluated an apparatus for separating tumor cells from a large volume of human marrow. This apparatus will permit preparation of large volumes of purged marrow for use in studies of intensive therapy with autologous marrow support. Bone marrow progenitor cell (CFU-GM) recovery following this IMP technique was 85% of the unpurged control, and suggests that marrow recovery following high dose systemic chemotherapy will not be adversely affected. A phase I study to evaluate marrow reconstitution following IMP is underway. Preliminary data suggest that this IMP method will not delay engraftment in breast cancer patients receiving high-dose chemotherapy and autologous bone marrow support, but further study is required.

Alterations in activity at auditory nuclei of the rat induced by exposure to microwave radiation: autoradiographic evidence using [14C]2-deoxy-D-glucose.

Autoradiographic maps of brain activity in rats exposed to pulsed or continuous-wave (CW) microwave radiation were made using [14C]2-deoxy-D-glucose ([14C]2-DG). Special emphasis was given to measurements of activity in the auditory system because previous work had shown that pulsed microwave radiation can elicit auditory responses in man and other animals. In particular, one middle ear was ablated in nine rats to attenuate the transmission of air-borne sound to one cochlea. The resulting imbalance in auditory input for four animals not exposed to microwave radiation was reflected as a bilateral asymmetry of [14C]2-DG uptake at the inferior colliculus and medial geniculate body. In contrast, a symmetrical pattern of uptake at these structures in an animal exposed to pulsed microwave radiation showed that this stimulus bypasses the middle ear in eliciting auditory responses. This result established the utility of the [14C]2-DG method for demonstrating a known effect of microwave radiation on brain activity. The results also revealed responses at auditory nuclei in 4 animals exposed to CW microwave radiation. These responses, which have not been observed with other methods, were evident at the power densities of 2.5 and 10 mW/sq. cm. To exclude the possibility that CW microwave radiation produced this result by direct action on brain tissue, additional data were obtained from two rats with one cochlea destroyed. In both animals, the uptake of [14C]2-DG at the inferior colliculus and medial geniculate body was virtually identical to the uptake in animals not exposed to microwave radiation, i.e. greatest on the side of the brain contralateral to the intact cochlea. This finding, coupled with the finding of a bilateral symmetry of [14C]2-DG uptake in the auditory pathways of animals with one middle ear ablated, confirmed the hypothesis that auditory responses to CW microwave radiation originate within the cochlea. Effects on brain activity outside of the auditory system were not found in qualitative analyses of autoradiographs for the conditions of exposure to CW microwave radiation noted above or for exposure to pulsed microwave radiation at the average power density of 2.5 mW/sq. cm.

A comparison using tissue electrical properties and temperature rise to determine relative absorption of microwave power in malignant tissue.

This paper compares two methods for determining the radio frequency absorbed power in tissue: from the measured electrical properties of the tissue, and from the induced temperature rise per unit time. In previous research, we measured the ratio frequency electrical properties of muscle, mammary gland, and malignant mammary tissue (SMT-2A mammary adenocarcinoma) in female W/Fu isogeneic rats. From those measurements we calculated for each tissue the power absorption versus frequency, and formed the ratio of malignant-to-normal power absorption. This ratio exhibited a peak within the 150 to 400 MHz range, indicating a selective absorption of power in this type of malignant tissue over that of the normal host tissue. In the present study, by an entirely different method, we have directly tested the results of our earlier research. We filled a 20-cm-long section of rigid coaxial line (ordinarily air filled) with either normal (beef muscle or fat) or malignant (SMT-2A) tissue, and measured the temperature increase versus time at the irradiated tissue surface for the same absorbed power in each tissue type. We made the measurements from 50 to 915 MHz, and found that the initial temperature increase per second per watt absorbed (dT/dt/Pa) was greater in malignant tissue than in muscle or fat at each frequency tested, with the greatest differences occurring below 450 MHz. Power absorption based on the measured values of dT/dt/Pa was again greatest for the malignant tissue (SMT-2A mammary adenocarcinoma) within the 150 to 400 MHz range.

The measured electrical properties of normal and malignant human tissues from 50 to 900 MHz.

The electrical conductivity and relative permittivity of malignant and normal human tissues were measured at frequencies from 50 to 900 MHz. The measurements were made between 23 and 25 degrees C using a network analyzer connected to a flat-ended coaxial probe that was pressed against the freshly excised tissue samples. The malignant tissues were of the following normal tissue origin: bladder, colon, kidney, liver, lung, lymph nodes, mammary gland, spleen, and testes. The normal tissues included: colon, kidney, liver, lung, mammary gland, and muscle. Normal tissue samples of bladder, lymph, spleen, and testes were not available. In general, at all frequencies tested, both conductivity and relative permittivity were greater in malignant tissue than in normal tissue of the same type. For tissues of the same type, the differences in electrical properties from normal to malignant were the least for kidney (about 6% and 4% average differences over the frequency range in permittivity and conductivity, respectively), and these differences were the greatest for mammary gland (about 233% and 577% average differences in permittivity and conductivity, respectively). To illustrate a potential use of these data in hyperthermia applications, frequency-selective heating of malignant tissue (modeled as a sphere) surrounded by host normal tissue is calculated from the measured electrical properties for certain tissues.

Heating patterns generated by phase modulation of a hexagonal array of interstitial antennas.

In this paper, we investigate an array of six interstitial microwave antennas used for hyperthermia cancer treatment. The purpose is to generate both uniform and controlled nonuniform heating patterns in biological tissue by phase modulating the signals applied to each antenna. The array consists of six antennas positioned on the corners of a hexagon. The distance between two diagonal antennas is 4 cm. The distributions of absorbed power per unit mass within the array are computed, and then converted into temperature distributions through a thermal conduction simulation. The SAR and temperature patterns are presented in both the lateral plane (perpendicular to the antennas) and the axial plane (parallel with the antennas). By proper phase modulation of microwave signals applied to each antenna, a uniform heating pattern can be produced within the entire array volume. Also, a peripheral heating pattern may be generated around the array; again, by using the proper phase modulation. The modulation schemes for generating both types of heating patterns are discussed.

Theoretical and measured electric field distributions within an annular phased array: consideration of source antennas.

The magnitude of E-field patterns generated by an annular array prototype device has been calculated and measured. Two models were used to describe the radiating sources: a simple linear dipole and a stripline antenna model. The stripline model includes detailed geometry of the actual antennas used in the prototype and an estimate of the antenna current based on microstrip transmission line theory. This more detailed model yields better agreement with the measured field patterns, reducing the rms discrepancy by a factor of about 6 (from approximately 23 to 4%) in the central region of interest where the SEM is within 25% of the maximum. We conclude that accurate modeling of source current distributions is important for determining SEM distributions associated with such heating devices.

Reception of microwaves by the brain.

The question of how pulsed microwaves induce auditory effects in animals and man leads to an examination of the parameters which determine the frequencies of maximum reception by the skull, and the positions of maximum energy deposition within the brain. The interaction of microwaves with a nerve membrane model and with live nerve cells is also discussed, as well as planned experiments to determine the microwave properties of membranes.

The effects of hyperthermia and hyperthermia plus microwaves on rat brain energy metabolism.

The effects of hyperthermia, alone and in conjunction with microwave exposure, on brain energetics were studied in anesthetized male Sprague-Dawley rats. The effect of temperature on adenosine triphosphate concentration [ATP] and creatine phosphate concentration [CP] was determined in the brains of rats that were maintained at 35.6, 37.0, 39.0, and 41.0 degrees C. At 37, 39, and 41 degrees C brain [ATP] and [CP] were down 6.0, 10.8, and 29.2%, and 19.6, 28.7, and 44%, respectively, from the 35.6 degrees C control concentrations. Exposure of the brain to 591-MHz radiation at 13.8 mW/cm2 for 0.5, 1.0, 3.0, and 5.0 min caused further decreases (below those observed for 30 degrees C hyperthermia only) of 16.0, 29.8, 22.5, and 12.3% in brain [ATP], and of 15.6, 25.1, 21.4, and 25.9% in brain [CP] after 0.5, 1.0, 3.0, and 5.0 min, respectively. Recording of brain reduced nicotinamide adenine dinucleotide (NADH) fluorescence before, during, and after microwave exposure showed an increase in NADH fluorescence during microwave exposure that returned to preexposure levels within 1 min postexposure. Continuous recording of brain temperatures during microwave exposures showed that brain temperature varied between -0.1 and +0.05 degrees C. Since the microwave exposures did not induce tissue hyperthermia, it is concluded that direct microwave interaction at the subcellular level is responsible for the observed decrease in [ATP] and [CP].

Power density, field intensity, and carrier frequency determinants of RF-energy-induced calcium-ion efflux from brain tissue.

To explain a carrier frequency dependence reported for radiofrequency (RF)-induced calcium-ion efflux from brain tissue, a chick-brain hemisphere bathed in buffer solution is modeled as a sphere within the uniform field of the incident electromagnetic wave. Calculations on a spherical model show that the average electric-field intensity within the sample remains the same at different carrier frequencies if the incident power density (Pi) is adjusted by an amount that compensates for the change in complex permittivity (epsilon *r) and the change of wavelength, as a function of carrier frequency. The resulting formula for transforming Pi is seen to follow the pattern of both positive and negative demonstrations of calcium-ion efflux that have been observed at carrier frequencies of 50, 147, and 450 MHz. Indeed, all results obtained at these three frequencies, when related by Pi's that produce the same average electric-field intensity within the sample, are seen to be in agreement; no prediction is contradicted by an experiment.

Design and analysis of annular antenna arrays with different reflectors.

The design and performance of annular antenna arrays with reflectors is presented. Arrays with three shapes of reflectors are analysed and simulated. These include the corner reflector, the circular reflector and the elliptical reflector. Power-density distributions within the annular arrays with and without reflectors are obtained by using the FDTD method. Also, the image theory method is used to verify the FDTD results in one case. By comparing the power-density distribution pattern of all four of the array designs (three with different reflectors, one without reflector), it is readily seen in each case that the array with reflectors yields better power-efficiency than the array without reflectors and the elliptical reflector yields the best performance. Comparisons of each array are made using 4, 6 and 8 antennae in the annular array. By using the optimized results of the elliptical reflector, the requirement for the input-power level to each antenna is greatly reduced.

Equalizing the electric field intensity within chick brain immersed in buffer solution at different carrier frequencies.

Presented here are the numerical relationships between incident power densities that produce the same average electric field intensity within a chick brain half immersed in buffered saline solution and exposed to a uniform electromagnetic field at carrier frequencies of 50, 147, and 450 MHz. Calculations are based on modeling the buffer solution as a spherical shell in air with an inner concentric sphere of brain tissue. The results support our earlier conclusion that calcium efflux results obtained at different carrier frequencies are in agreement when related by the electric field within the brain.

Characteristics of transverse electric and magnetic field transmission cells at extremely low frequencies.

Transverse electric and magnetic field (TEM) cells are often designed to subject samples to electromagnetic radiation of intrinsic impedance (E/H) that is the same as in free space, 377 omega. Earlier work has shown this value to be correct for the RF region above about 2 kHz. In this study, measurements of magnetic fields in the extremely low frequency regions and at DC indicate the E/H ratio to be around 300 omega for frequencies less than 2 kHz in cells of a particular design. This lower value indicates that care should be taken in estimating AC magnetic field intensities from electric field measurements in TEM cells at frequencies below 2 kHz.