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Three square (L, M, MS) and one rectangular (HN) applicators with captive boluses were provided by the Clini-Therm Corporation for evaluation. Surface cooling is achieved by attaching a mineral oil captive bolus to the built in water-circulating tubes at the aperture of the applicators. These applicators were tested on a phantom with a 2-cm fat slab over 10-cm-thick muscle. Surface and sagittal heating patterns were obtained using a thermograph. All captive-bolus applicators have heating patterns similar to that of the regular Clini-Therm applicators. Due to hot spots at the edges of the applicators where the E fields terminate, these modified applicators should not be placed in direct contact with patients when boluses are not used. Tests with Clini-Therm regular water bolus instead of the captive oil bolus indicated that the orientation of water flow should be parallel to the E field to minimize perturbation of the heating patterns. Thermal conduction studies showed that the captive bolus reacts too slowly for skin temperature control. The modified captive bolus applicators did not improve the performance of the system.

A rare site for an epidermoid cyst is herein reported. A 40-year-old female was admitted for treatment of a right plantar tumor. The tumor was multiloculated, soft, nodular and in the range of about seven by seven centimeters. We initially diagnosed it as a saccular hemangioma, but it was finally diagnosed as an epidermoid cyst with foreign body reaction from the surgical specimen. Epidermoid cysts can occur in various parts of the body. Trauma is regarded to be the chief cause of them. Due to their various locations and appearance, they are sometimes difficult to diagnose. In such cases, however, it is generally agreed that pathological study is mandatory for a correct diagnosis.

The specific absorption rate (SAR), which is the mass-normalized rate of energy absorption by a biological body, has been used by hyperthermia researchers to characterize energy deposition and heating patterns in tissues and in biological models. Before thermal diffusion and blood flow alter the temperature distribution, energy deposition and heating rate (HR) are related by constants. Therefore SAR is usually calculated from the initial rate of temperature rise measured during heating. SAR is an appropriate parameter for theoretical calculations or electric field measurement in tissue. However, the ultimate parameter in hyperthermia is temperature. Instead of computing the temperature rise from SAR (W/kg) and specific heat (kcal/kg.degree C) which were originally obtained from temperature measurements, it is simpler and more convenient to use HR (degree C/W-min) directly, especially when heterogeneous tissues are involved. The advantages of using HR instead of SAR in hyperthermia clinics are discussed.

Insulin has both short- and long-term effects on cellular metabolism. The short-term effects are known to involve the insulin receptor, a protein kinase capable of phosphorylating itself and other proteins. The role of the receptor was elucidated by studies of a mutant insulin receptor which lacked kinase activity and inhibited several actions of insulin. The long-term effects of insulin could be demonstrated by its growth-promoting effect on hepatoma cells, and by the suppression in transfected hepatoma cells of hepatitis B virus antigen production in a dose-dependent manner. The process whereby insulin appears to regulate gene expression is not clearly understood.

Complementary DNA of a rat brain glucose transporter gene was used to examine the expression of glucose-transporter messenger RNA in paired human hepatocellular carcinomas and adjacent nontumorous liver tissues, as well as in human hepatoma cell lines and human fetal liver samples. High expression of a major 2.8-kilobase glucose-transporter transcript was seen in all hepatoma cell lines and fetal liver samples examined, whereas a much lower level of expression was observed in liver tissues. When pairs of liver tissues were examined, elevation of glucose-transporter RNA levels was observed in most of the hepatocellular carcinoma tissues examined.

Endothelin is a vasoconstrictor peptide which has recently been isolated and sequenced from the vascular endothelial cells. It was reported to increase blood pressure in vivo and produce a prolonged contraction with a slow onset in vitro. The purpose of this study was to investigate whether endothelin can lower the portal pressure as another endogenous vasoconstriction peptidevasopressin (AVP) can. Heart rate, systemic blood pressure, portal pressure, and portal vein blood flow were measured. Effects of endothelin on these parameters were compared with those of AVP. Endothelin 10(-10) mol/Kg significantly decreased all of the parameters mentioned. At the higher dose (5 x 10(-10) mol/Kg), however, the portal pressure and blood pressure were increased and portal vein blood flow was unchanged. On the other hand, AVP decreased the portal pressure and portal vein blood flow but elevated the systemic blood pressure. In vitro experiments revealed that endothelin contracted both tail artery and portal vein of rat and vasopressin contracted only tail artery. We concluded that although both are endogenous vasoconstricting peptides, endothelin and AVP affect differently on arterial and venous vascular beds as well as on portal pressure.

Human hepatoma cell lines were studied for the expression of platelet-derived growth factor (PDGF), insulin-like growth factor-I (IGF-I) and their receptors at the mRNA level. Transcripts of PDGF were consistently detected in these cell lines. In addition, some cell lines also expressed PDGF receptor RNA. Moreover, RNA of IGF-I and its receptor were detected in every cell line examined. These results suggest that autocrine regulation may be an important mechanism for the maintenance of the transformed state of human hepatoma cells.

A rapid, precise, and accurate liquid chromatographic (LC) method is described for the determination of benzoyl peroxide (BP) in acne preparations. BP is extracted from a water dispersion of the preparation with dichloromethane (DCM), and an aliquot is eluted from a C-18 reverse phase LC column with acetonitrile-0.10 M aqueous NaClO4. Selective and sensitive quantitation is accomplished with a reductive mode electrochemical detector. This detector is an order of magnitude more sensitive than a 240 nm UV absorption detector; the lower limit of detection is 2 ng for a 4 microL injection. The recovery of BP is 99.4% and the detector response is linear to at least 2 micrograms per 4 microL injection.

The expression of 20 known cellular proto-oncogenes in human well-differentiated hepatoma cell line Hep3B and poorly-differentiated hepatoma cell line HA22T/VGH was studied by Northern blot hybridization. Among the cellular proto-oncogenes examined, both cell lines express protein kinase genes including fps, mos and raf; PDGF B chain sis gene; GTP/GDP binding protein gene Ha-ras and nuclear protein genes including fos and myc. The expression of yes, abl, ros, src, erb-B, erb-A, fms, Ki-ras, myb, rel and bas genes was not detected in both cell lines.

Insulin-like growth factor (IGF) I and IGF-II were measured by radioimmunoassay in the sera of seven patients with acromegaly, 36 normal control subjects, 15 patients with chronic hepatitis, 15 patients with cirrhosis, 25 patients with hepatocellular carcinomas (HCCs) who did not have hypoglycemia, 20 patients with HCCs who did have hypoglycemia, and 10 patients with metastatic liver tumors. Both IGF-I and IGF-II levels decreased as liver disease progressed from the normal control stage to chronic hepatitis and cirrhosis, and both levels reflected the severity of liver disease. Patients with HCCs who had hypoglycemia had relatively higher IGF-II levels in their sera in comparison with those who did not have hypoglycemia (272 +/- 167.5 ng/ml vs 110.4 +/- 85.9 ng/ml [mean +/- SD], p less than 0.0005), despite the fact that those with hypoglycemia had more advanced liver cancer and had lower IGF-I levels in sera (16.7 +/- 14.1 ng/ml vs 46.8 +/- 47.9 ng/ml, p less than 0.002). It is possible that a labile IGF-II material is produced by the cancer cells of patients with hypoglycemia. This factor is reactive to the IGF-II receptor and partially cross-reacts with an antibody to IGF-II; it accounted for the mildly elevated levels of serum IGF-II. Hypoglycemia may be an integral effect of relatively elevated IGF-II like material and an advanced liver cancer. Also, higher serum alpha-fetoprotein (AFP) levels were more frequently found in patients with hypoglycemia who had relatively elevated IGF-II levels and short survivals.

In previous research, we found that sodium-dependent high-affinity choline uptake in the hippocampus and frontal cortex of the rat was lowered after acute (45 min) exposure to low-level 2450-MHz pulsed microwaves (power density 1 mW/cm2; average whole body specific absorption rate, 0.6 W/kg; 2 mu sec pulses, 500 pps). In the present experiment, we investigated developments of tolerance and classical conditioning to these effects of microwaves. Rats were exposed to microwaves in cylindrical waveguides in 10 daily sessions (45 min per session). In an 11th session, we subjected the rats to either microwave (study of tolerance) or sham exposure (study of conditioned effect) for 45 min, and immediately measured choline uptake in the hippocampus and frontal cortex. We found that tolerance, a decrease in response to microwaves, developed to the effect of microwaves on choline uptake in the hippocampus, but not in the frontal cortex. Conditioned effects were also observed: an increase in choline uptake in the hippocampus and a decrease in uptake in the frontal cortex. These data suggest that the effects of microwaves on choline uptake in the hippocampus and frontal cortex are classically conditionable, probably to cues in the exposure environment.

Microwave irradiation of rats by circularly polarized, 2,450-MHz, pulsed waves (2-microseconds pulses; 500 pps) was performed in waveguides to determine effects on ethanol-induced hypothermia and on ethanol consumption. Rats injected intraperitoneally with ethanol (3 g/kg in a 25% v/v water solution) immediately after 45 min of microwave irradiation exhibited attenuation of the initial rate of fall in body temperature, which was elicited by the ethanol, but exhibited no significant difference in maximal hypothermia as compared with that of sham-irradiated rats. Microwave irradiation did not affect the consumption of a 10% sucrose (w/v) solution by water-deprived rats. However, it enhanced the consumption of a solution of 10% sucrose (w/v) + 15% ethanol (v/v) by water-deprived animals. These results were obtained at a specific absorption rate (SAR) of 0.6 W/kg, which rate of energy dosing would require a power density of 3-6 mW/cm2 if exposure of the animals had occurred to a 12-cm plane wave.

The changes in heating patterns of interstitial microwave antennas at different insertion depths were investigated in a static phantom at 915 MHz. Antennas for the Clini-Therm Mark VI system were inserted 5-15 cm into muscle-equivalent material, through nylon catheters. The phantom was heated with arrays of antennas at 2 cm spacings for 60 s at 15 W per antenna. Midplane and transverse heating patterns were determined thermographically with the antennas inserted parallel or perpendicular to the split of the phantom. Hot spots, independent of heating near the junction plane, were observed in the midplane of the 2 x 2 and 2 x 4 arrays at 2.8 cm from the insertion end. The magnitudes of these hot spots were reduced by 40-45 per cent as insertion depth was increased from 7 to 10.5 cm. With insertion depths of more than 11.5 cm the hot spots gradually diminished and heating occurred mostly near the junction plane. The observed heating patterns were caused by changes in impedance of the antenna arrays at different insertion depths. The impedance mismatch had resulted in different wave propagation within the tissue material which produced different radiation patterns. During treatments, because heating varies with insertion depth, great care must be exercised in monitoring temperatures.

To study the expression and the regulation of hepatocyte markers, we have undertaken to establish human hepatoma cell lines of various phenotypes. We now report the establishment of a new human hepatoma cell line, HA22T/VGH. This cell line has many of the properties of human hepatocellular carcinoma. Only 5 of 15 plasma proteins investigated were detected in the medium of a 10-day-old HA22T/VGH culture. However, when the HA22T/VGH cells and a clonal derivative, C5, were cultured in an aggregated form, all 15 plasma proteins were found in the culture medium. These results indicate that hepatoma cell lines with different phenotypes can be established, and they provide a good experimental framework to investigate differentiation of human hepatocytes.

In human hepatoma cell line Hep3B/T2, the human heat-shock-inducible gene hsp70 could be induced by insulin. The dose-dependent insulin effect correlates very well with the dissociation constant of the insulin receptor, indicating that the insulin effect is mediated by the insulin receptor. The expression of hsp70 gene was neither significantly induced by growth factors of epidermal and platelet-derived growth factors, nor by tumor promoter, calcium ionophore, cAMP, and glucocorticoids. These results indicate that the induction of expression of hsp70 gene by insulin is very specific and not cell cycle-related. Furthermore, the insulin-induced expression of hsp70 gene is transient, occurring specifically from 4 to 8 h after insulin addition.

Human growth hormone (HGH) was extracted from acromegalic pituitary tumors at pH 10.5 and precipitated with ammonium sulfate at 20-40% saturation. It was purified on a Sephadex G-100 column to yield monomeric HGH. The tumor-HGH was indistinguishable from the authentic one in polyacrylamide gel electrophoresis at pH 8.3 or in the presence of sodium dodecyl sulfate, high-performance liquid chromatography, radioimmunoassay, peptide map, amino acid composition and N-terminal partial amino acid sequence. The tumor-HGH is active in the tibia assay and bodyweight gain test in hypophysectomized rats with comparable potency to that of the authentic sample.

Hearts from 81 frogs were divided into 10 groups and placed individually in a temperature-controlled waveguide filled with Ringer's solution. Hearts of Group A served as controls. Hearts of the other nine groups were treated with pulsed microwaves (2450 MHz, 10 microseconds, 100 pps) that ranged from a specific-absorption rate (SAR) of 2 to 200 W/kg. In several groups, an electrocardiogram was used to synchronize microwave irradiation with the heartbeat. In one group of frog hearts exposed at 200 W/kg, the heart rate slowed down sharply 25 minutes after irradiation, by which time the temperature of the Ringer's solution had risen by 7 degrees C. In another group exposed at 200 W/kg but under constant temperature, the heartbeat almost paralleled that of the control group. Other groups, including exposures of frog hearts to 16 Hz modulated CW or pulsed fields at SARs from 3 mW/kg to 8.55 W/kg, showed no effects of pulsed microwave irradiation on heartbeats; no brachycardia or tachycardia were found at constant temperatures.

In a series of experiments, we investigated the effects of pulsed low-level microwave irradiation on amphetamine-induced hyperthermia in the rat. Rats were irradiated in a 2,450-MHz cylindrical waveguide exposure system at 1 mW/cm2, 2 mus pulses, 500 pps, average SAR of 0.6 W/kg. Acute (45 min) exposure to microwaves attenuated amphetamine-induced hyperthermia. This effect was blocked by pretreatment of the animals with the narcotic antagonist naloxone. In another experiment, rats were subjected to ten daily sessions of microwave exposure (45 min/session). On day 11, amphetamine-induced hyperthermia was studied in the animals immediately after a session of either microwave or sham exposure. Similar to the acute effect, amphetamine-induced hyperthermia was attenuated in rats irradiated with microwaves (unconditioned effect). In the sham-irradiated animals we observed a potentiation of the amphetamine-induced hyperthermia, which was a conditioned effect of microwaves. Thus, the conditioned effect (potentiation) was opposite in direction to the unconditioned effect (attenuation). No tolerance developed to the unconditioned effect after subchronic exposure. Furthermore, both conditioned and unconditioned effects of microwaves on amphetamine-induced hyperthermia could be blocked by treatment with naloxone. These data suggest that microwave irradiation may activate endogenous opioids, which in turn alter the actions of psychoactive drugs, and the effect of microwaves on drug action can be classically conditioned.

Previous reports have shown that microwave exposure can decrease the beating rate of isolated rat hearts. These experiments were conducted at room temperature and with the hearts exposed to air. We observed arrhythmia frequently at room temperature, and the variation of heart beat was so large that it makes the results difficult to reproduce. Therefore, we employed a double-circulating system to provide perfusion through the coronary artery and around the outside of the heart to maintain the rat hearts at 37.7 degrees C. No arrhythmias were observed in our experiments, and the hearts were beating for at least 1 h. The effects of 16-Hz modulated 2,450-MHz pulsed microwaves (10 microseconds, 100 pps) on the beating rate of 50 isolated rat hearts were studied. Results showed no statistically significant changes of heart rate in exposed groups at SARs of 2 and 10 W/kg compared with the control group. The effect seen at 200 W/kg was shown to be similar to that resulting from heating the heart.

One hundred and two isolated frog hearts were divided into ten groups and placed individually in a waveguide filled with Ringer's solution and exposed to 2,450-MHz CW radiation at 2 and 8.55 W/kg. Heart rate was recorded using one of the following methods: 3-M KCl glass electrode, ultrasound probe, tension transducer, Ringer's solution glass electrode, and a metal wire inserted in the Ringer's solution electrode. An accelerated decrease of heart rate was observed only in those groups recorded using the 3-M KCl electrode and the metal wire Ringer's solution electrode. No effect was found in the other groups. These results indicate that bradycardia in isolated hearts could be caused by electrode artifacts resulting from the intensification of electromagnetic fields.

Spheres composed of phantom muscle of radius 1.5, 2, 3, 4, 6 and 8 cm, as well as birds (parakeets, quail, pigeons, chickens, turkeys) were exposed to far-field plane waves at power densities of incident radiation between 182 and 560 mW/cm2 and at frequencies of 775, 915 and 2450 MHz. Specific absorption rate (SAR) patterns were determined by thermographic techniques for both spheres and birds. The measured SAR patterns in spheres were comparable to those from theoretical predictions. The SAR patterns in birds, however, varied markedly from those obtained from spheres of comparable mass. The results indicate that the geometrically complex animal is not represented by simple geometric models for making absorption studies. Thermograms of birds exposed in the flying position indicated that the SAR is high in the wings. The behavioral response of the birds to the exposure was variable. Threshold power densities for biological or behavioral reactions were determined for each bird at all three frequencies. The lowest power density associated with reactivity by the chicken was 5.8 mW/cm2 (corresponding to SARs of 3.1 W/kg in the head and 3.9 W/kg in the neck) at 775 MHz.

We have grown a human hepatoma cell line, designated as HA22T/VGH, from a 52-yr-old male hepatoma patient since July 1, 1980. This cell line has been subcultured more than 100 passages. The chromosome analysis of HA22T/VGH indicated that the chromosome numbers varied from 70 to 146, with the mode of 73. Methylcellulose soft agar assay showed that approximately 40% of the HA22T/VGH cells formed colonies. The HA22T/VGH produced tumors in nude mice. Histopathological studies of the tumor revealed the arrangement of hepatoma. Detected by the complement fixation method HA22T/VGH cells secreted ceruloplasmin, Factor B, C3, C4, Gc-globulin and alpha 1-acid-glycoprotein. These cells contained the liver associated enzymes: alanine amino transferase, tyrosine amino transferase and gamma-glutamyl transferase. HBsAg and alpha-fetoprotein were not detectable in the HA22T/VGH culture media or cell lysates by the radioimmunoassay.

The dielectric properties of various test samples of phantom tissue were measured using an automated and temperature-controlled slotted line. The ingredients for phantom materials were determined for simulating high-water content tissue at 13.56, 27.12, 40.68, 70, 100, 200, 300, 433, 750, 915, and 2,450 MHz. The ingredients consisted of water, TX-150 (a gelling agent), sodium chloride, and polyethylene powder (200-2,450 MHz) or aluminum powder (13.56-100 MHz). The dielectric constant and conductivity of these materials at different temperatures (15, 22, 30 degrees C) were characterized.

Transfection of human hepatoma cell lines with cloned HBV DNA resulted in the secretion of large amounts of hepatitis B surface antigen (HBsAg) and core-related antigens (HBc/HBeAg) if well-differentiated cell lines were employed. Synthesis of both viral antigens was the highest in cell line HuH-7 and continued for approximately 25 days. Particles resembling hepatitis B virions (Dane particles) by morphology, density and by the presence of the preS1 surface antigen were released from the transfected HuH-7 cells into the culture medium. These particles produced in vitro were also indistinguishable from the naturally occurring hepatitis B virions in containing the virus-associated DNA polymerase and mature HBV genomes. Restriction analysis of these DNA molecules was compatible with the nucleotide sequence of the transfecting HBV DNA sequence. Viral surface antigens and core proteins present in the culture medium were fractionated and characterized by immunoprecipitation and SDS--PAGE after labeling with [35S]methionine. Antisera specific for X-gene products identified in cell extracts two hitherto unknown HBV gene products. This system thus provides a new approach to open questions regarding HBV-related gene function and HBV replication.

Using complementary DNAs of human insulin-like growth factors as probes, expressions of the insulin-like growth factors I and II mRNA were examined in seven human hepatoma tissues and their adjacent nontumorous livers. The level of insulin-like growth factor I mRNA in hepatoma was lower than that in the nontumorous liver control. This phenomenon was probably caused by the low expression of human growth hormone receptor in hepatoma tissues. The levels of insulin-like growth factor II mRNA vary among hepatomas. Some show elevated expression; some have diminished expression compared to their nontumorous liver counterparts. In four of the seven hepatomas, expression of fetal forms of insulin-like growth factor II transcripts was observed and may represent dedifferentiation of insulin-like growth factor II expression during hepatocarcinogenesis.

Both positive and negative biological effects of microwaves on drug actions in rats exposed to 1-mW/cm2, 2,450-MHz microwaves have been reported by several investigators. We conducted dosimetry studies for seven different exposure conditions to determine whether these different results could be due to the rats having been exposed differently. They included anterior and posterior exposures in a circular waveguide, near field, far field with E- or H-field parallel to the long axis of the body and dorsal exposure in a miniature anechoic chamber with E- or H-field parallel to the long axis of the body. The average specific absorption rates (SARs) in the head, tail, and body of the exposed rats were measured by means of a calorimetry system. The local SARs at eight locations in the brain were determined by temperature measurement with Vitek probes. Intensive coupling of energy to the tail when it was exposed parallel to the E-field was shown by thermography. For the same average incident power density, the average SARs in the heads of rats were about two times higher in the circular waveguide than for other exposures. The local SARs in the brain varied for different exposure conditions. Statistical comparisons of SARs under the different exposure conditions are presented.

We have previously demonstrated that the human insulin receptor, mutated in the ATP-binding domain of the beta-subunit, is kinase-defective and fails to mediate multiple post-receptor actions of insulin in stably transfected Chinese hamster ovary cells (Chou, C.-K., Dull, T. J., Russell, D. S., Gherzi, R., Lebwohl, D., Ullrich, A., and Rosen, O. M. (1987) J. Biol. Chem. 262, 1842-1847). This study addresses the role of protein-tyrosine kinase activity in insulin-mediated receptor down-regulation. Although the mutant insulin proreceptor was properly processed and able to bind insulin like the wild-type human receptor, it differed from the latter in the following respects: 1) it failed to mediate internalization of surface-bound radiolabeled ligand; 2) it did not undergo short- or long-term down-regulation in response to 1 microM insulin; 3) it did not exhibit ligand-promoted receptor turnover; and 4) it was not phosphorylated on either tyrosine or serine residues in response to insulin. Although the cells transfected with the mutant receptor failed to respond to insulin-mediated insulin receptor down-regulation, they were able to down-regulate their insulin-like growth factor I receptors in response to insulin-like growth factor I or high concentrations of insulin and were sensitive to monoclonal antibody-induced down-regulation of their insulin receptors. Antibody-mediated receptor internalization alone, however, was unable to mimic at least one action of insulin, thymidine incorporation into DNA, and did not lead to any phosphorylation of the receptor. It is concluded that either the protein-tyrosine kinase activity of the insulin receptor or its phosphorylation state is essential for ligand-mediated receptor down-regulation.

Microwave hyperthermia applicators are generally characterized at two or three frequencies. These frequencies usually are at the extremes of the applicator's operating range. However, such applicators are often used at arbitrary frequencies within their range. The most common reason for choice of frequency is to achieve acceptable coupling in the clinical configuration with a given patient. Occasionally a spurious transmission pattern will result from the frequency chosen that can lead to undetected high power densities well away from the target volume. In the present report, such a situation is discussed. The transmission pattern resembled "horns" reported at the field edge of certain accelerators. This pattern resulted in the inability to heat the target volume and the production of thermal blisters outside that volume. Such an occurrence underscores the need to characterize the transmission characteristics of a microwave hyperthermia applicator at all the frequencies used.

The cytosolic fractions from epidermal growth factor (EGF)-treated A431 cells exhibit a marked increase in activities of ATP.Mg-dependent protein phosphatase and its activating factor (protein kinase FA) when compared to controls in the absence of EGF. By contrast, the Triton X-100-solubilized membrane fractions from the same EGF-treated cells exhibit a corresponding decrease in protein kinase FA activity. The EGF-dependent activation of protein kinase FA and ATP.Mg-dependent protein phosphatase occurred within physiological concentrations of EGF (ED50 = 5 x 10(-10) M). The changes of kinase and phosphatase activities which were measured concomitantly exhibit very similar characteristics as to EGF sensitivity and time dependence. The EGF-induced kinase and phosphatase activation occurred very rapidly, reaching the maximal activity levels within 3 min. Moreover, the EGF effect is transient; both EGF-stimulated phosphatase and kinase activities returned to control levels within 30 min. Taken together, the results suggest that EGF may induce the activation of kinase FA in the membrane and thereby promotes the activation of ATP.Mg-dependent phosphatase in the cytosol. Exposure of A431 cells to exogenous phospholipase C also resulted in the activation of endogenous kinase FA and ATP.Mg-dependent phosphatase in a similar pattern produced by EGF. This further suggests that phospholipase C can mimic EGF to mediate the activation of kinase FA and ATP.Mg-dependent phosphatase in A431 cells. By its dual role as a multisubstrate protein kinase and as an activating factor of multisubstrate protein phosphatase, protein kinase FA may represent a transmembrane signal of EGF.

The contribution of the ossicles (middle-ear bones) to auditory perception of microwaves was evaluated by the brain-stem evoked response (BER). Amplitude and latency of BERs were recorded from guinea pigs that were stimulated at various intensities by acoustic pulses coupled to the auditory canal or via bone conduction, and by microwave pulses. Blocking of the external ear, middle-ear damping, and middle-ear destruction produced little change in the BERs that were elicited by microwave pulses. Results indicate that activity in the central auditory pathway as induced by pulsed microwaves only requires stimulation of the cochlea. Conduction of pressure waves through the bones of the calvarium appears to be the mechanism responsible in perception of pulsed microwaves.

The effects of various psychoactive drugs were studied in rats exposed for 45 min in a circularly polarized, pulsed microwave field (2450 MHz; SAR 0.6 W/kg; 2-microseconds pulses, 500 pps). Apomorphine-induced hypothermia and stereotypy were enhanced by irradiation. Amphetamine-induced hyperthermia was attenuated while stereotypy was unaffected. Morphine-induced catalepsy and lethality were enhanced by irradiation at certain dosages of the drug. Since these drugs have different modes of action on central neural mechanisms and the effects of microwaves depend on the particular drug studied, these results show the complex nature of the effect of microwave irradiation on brain functions.

The thermographic method for determining specific absorption rate (SAR) in animals and models of tissues or bodies exposed to electromagnetic fields was applied to the problem of quantifying the current distribution in homogeneous bodies of arbitrary shape exposed to 60-Hz electric fields. The 60-Hz field exposures were simulated by exposing scale models of high electrical conductivity to 57.3-MHz VHF fields of high strength in a large 3.66 X 3.66 X 2.44-m TE101 mode resonant cavity. After exposure periods of 2-30 s, the models were quickly disassembled so that the temperature distribution (maximum value up to 7 degrees C) along internal cross-sectional planes of the model could be recorded thermographically. The SAR, W', calculated from the temperature changes at any point in the scale model was used to determine the SAR, W, for a full-scale model exposed to a 60-Hz electric field of the same strength by the relation W = (60/f')2 . (sigma'/sigma) . W' where f' is the model exposure frequency, sigma' is the conductivity of the scale model at the VHF exposure frequency, and sigma is the conductivity of the full-scale subject at 60 Hz. The SAR was used to calculate either the electric field strength or the current density for the full-scale subject. The models were used to simulate the exposure of the full-scale subject located either in free space or in contact with a conducting ground plane. Measurements made on a number of spheroidal models with axial ratios from 1 to 10 and conductivity from 1 to 10 s/m agreed well with theoretical predictions. Maximum current densities of 200nA/cm2 predicted in the ankles of man models and 50 nA/cm2 predicted in the legs of pig models exposed to 60-Hz fields at 1kV/m agreed well with independent measurements on full-scale models.

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 less than .01) decrease only of food consumption during the 5-mW/cm2 exposure; other variables were not significantly different between exposed and control groups.

The human hepatoma Hep3B cells contain integrated hepatitis B viral genome and continually secret hepatitis B surface antigen (HBsAg). The production of HBsAg (but not alpha-fetoprotein) was suppressed by addition of low concentrations (0.1-1 nM) of insulin into serum-free medium. In addition, the suppression of HBsAg production by insulin was paralleled with the decrease in HBsAg mRNA abundance. Insulin also cause a rapid rate of disappearance of HBsAg mRNA (t 1/2, 2 h) in Hep3B cells. The Hep3B cells carry specific receptor with high affinity for insulin (Kd = 1.8 nM). The receptor showed an insulin-dependent protein tyrosine kinase activity. The half-maximal insulin concentration for the activation of the receptor kinase was about 5 nM. Only very high concentrations of insulin-like growth factor I and human proinsulin can compete for the insulin receptor binding and suppress HBsAg production, this suggests that insulin may act through its receptor binding to suppress HBsAg expression in human hepatoma Hep3B cells.

The specific absorption rate (SAR) patterns of two 434-MHz hyperthermia applicators, models TCA 434-1 (9 X 20 cm) and TCA 434-2 (13 X 25 cm), were evaluated thermographically using a phantom model. The phantom model consisted of a 2-cm-thick layer of fat and a 10-cm depth of muscle contained in a 30 X 30 cm base Plexiglas box. The model was bisected in the middle. Polyester screens at the interface allowed the synthetic gel to make electrical contact between the two halves of the muscle tissue. Octyl alcohol was applied to the fat interface to ensure continuity of dielectric properties. Thermograms were taken for both applicators over the following areas of the exposed model: (1) fat surface, (2) internal surface with E-field parallel to interface, and (3) internal surface with E-field perpendicular to interface. SAR's were calculated from the temperature rise (8 degrees C maximum), net input power (550-650 W), exposure time (15-60 s), and specific heat of the muscle (0.86 kcal/kg degrees C). A factor of 0.42 needs to be multiplied to correct for the specific heat of fat. High localized SAR's along the broad sides of the applicators were seen when the applicators were in direct contact with the phantom. With the use of a 0.8-cm polystyrene foam spacing, the SAR's within the aperture of the applicators were relatively uniform. The patterns of the two applicators were quite similar. However, the TCA 434-1 applicator is smaller and more applicable for clinical conditions.

Conventional metal electrodes can cause serious modification and intensification of the rate of energy absorption in tissues during exposure of animals to microwaves. Carbon-loaded Teflon with a conductivity close to that of tissue has been implanted and maintained for four to six months at cortical and subcortical locations in rabbits. The EEG and its spectrum as recorded from the carbon-loaded Teflon electrodes are comparable to those recorded from conventional metal electrodes. Histological examination showed good tissue compatibility. Recordings made during acute exposure of rabbits to microwave radiation (2450-MHz) at 100 mW/cm2 showed no electromagnetic interference. The results indicate that carbon-loaded Teflon electrodes can be implanted chronically to record the EEG in animals during the course of microwave radiation.

The construction of a modified coaxial cable as an intracavitary microwave applicator suitable for use in some vaginal and rectal cancers is presented. Thermometry was performed for microwave frequencies of 300, 400, 650, and 915 MHz. Temperature profiles in tissue phantoms were obtained with non-perturbing temperature probes and thermography, and the data were compared with those obtained in dogs. The temperature profiles were dependent on the frequency of the microwaves and the insertion depth of the applicator. In addition, an acrylic cylindrical spacer external to the applicator also altered the heating pattern. Therefore, with proper combinations of frequency, insertion depth and spacer, the applicator can be used for heating tumors in some clinical situations.

Hep 3B, a human hepatoma cell line was examined for its RNA hybridizable to the hepatitis B virus sequence. Using probes that covered different regions of the hepatitis B virus genome, five species of RNA were observed of sizes 4.0, 3.3, 2.9, 2.6 and 2.2 kilobases. The RNAs covered surface antigen gene, pre-S and X regions. None of them had a core antigen sequence. RNA with a 4.0 kilobase size was the most abundant. Using S1 nuclease analysis, its 5' end of hepatitis B virus sequence was mapped at pre-S region and its 3' end of viral sequence was mapped at DR region.

The effects of somatostatin (SRIF), insulin, and triiodothyronine (T3) on the growth of human hepatoma cells were investigated on the well-differentiated human hepatoma cell line Hep3B. Results showed that both insulin and T3 can stimulate cell growth of serum starved Hep3B cells at physiological concentrations. SRIF alone showed little growth-promoting activity. When added concurrently with insulin, however, SRIF suppressed the insulin-induced cell proliferation in a dose-dependent manner. On the other hand, SRIF had no inhibitory effect on T3-induced cell proliferation. SRIF is labile in the medium, with a half-life of about 2 h during culture incubation. SRIF did not disturb the insulin binding to its surface receptors nor inhibit the insulin-dependent receptor kinase activity of Hep3B cells in vitro. These results suggest that postreceptor regulation may be involved. The selective suppression by SRIF of insulin-induced cell growth provides an unique approach to the study of insulin actions on proliferation of human hepatoma cells.

Tissue-substitute models consisting of layers of synthetic, electrically equivalent subcutaneous fat, muscle, and bone shaped in conformation with the normal anatomy are used for rapid determination of distribution of temperature and specific absorption rate throughout the tissues when exposed to electromagnetic radiation. The surfaces of the bisected models are approximated during a short exposure period, then separated and scanned with a thermograph. A method was developed to eliminate the electrical discontinuity at the bisected surfaces while allowing separation and subsequent thermographic scanning. A thin layer of silk screen wetted with propylene glycol saturated with sodium chloride was used at the fat interface and a 0.9% sodium chloride solution was used to wet the screen at the muscle interface to eliminate electrical discontinuity during exposure to 27.12-MHz diathermy. Tests showed that in the presence of an electrical discontinuity the heating pattern was grossly distorted. With the method used, the electrical discontinuity is minimized and the subsequent thermographic scanning reveals that the heating pattern is equivalent to that of an intact model.

The level of c-myc transcript was examined in liver samples from seven hepatoma patients. Transcripts were detected in all the normal liver parts examined; in contrast, in two hepatoma parts, there was a dramatic reduction in c-myc transcripts. The restriction enzyme pattern of c-myc gene appeared the same among samples. The data suggest that c-myc gene expression might not be required for the maintenance of the tumor state in human liver carcinogenesis.

The effect of microwave irradiation on naloxone-induced withdrawal syndrome was studied in rats made morphine dependent by subcutaneous implantation of morphine pellets. Morphine-dependent rats were either exposed to pulsed low-level microwaves (2450 MHz, 1 mW/cm2, 500 pps, 2 musec pulses) or sham-irradiated for 45 min before the naloxone injection. We found that microwave-exposed rats showed significantly less weg-dog-shakes and had higher body temperature than the sham-exposed animals during withdrawal. There was no significant difference in the incident of diarrhea between the two groups of animals. These data further support the results of our previous research suggesting that pulsed low-level microwave irradiation activates endogenous opioids in the rat.

Two monoclonal antibodies, designated 4C4 and 4G1, were produced by immunization of BALB/c mice with a human esophageal carcinoma cell line, CE69T/VGH, followed by fusion of the spleen cells from an immunized mouse with myeloma cells NS-1. 4C4 showed strong binding activity to three human esophageal carcinoma cell lines and one human hepatoma cell line, but not to any other cell lines tested. 4G1 reacted with three human esophageal carcinoma cell lines and four other cell lines. By peroxidase-antiperoxidase staining, 4C4 and 4G1 detected antigens of the epithelial cells on 10 pairs of esophageal carcinoma and normal esophageal specimens. 4G1 recognized a CE69T/VGH antigen with a molecular weight of 180K. Since 4G1 also reacted with purified carcinoembryonic antigen (CEA) and immunoprecipitated 125I-CEA, 4G1 seems to be an antibody recognizing CEA produced by CE69T/VGH cells. Since 4C4 also bound to the epithelial cells of normal uterine, vaginal, breast and liver tissues, it seems to recognize an epithelial antigen, and can be used to characterize the antigen in the specialization or differentiation of epithelial cells.

Sodium-dependent high-affinity choline uptake was measured in various regions of the brains of rats irradiated for 45 min with either pulsed or continuous-wave low-level microwaves (2,450 MHz; power density, 1 mW/cm2; average whole-body specific absorption rate, 0.6 W/kg). Pulsed microwave irradiation (2-microseconds pulses, 500 pulses/s) decreased choline uptake in the hippocampus and frontal cortex but had no significant effect on the hypothalamus, striatum, and inferior colliculus. Pretreatment with a narcotic antagonist (naloxone or naltrexone; 1 mg/kg i.p.) blocked the effect of pulsed microwaves on hippocampal choline uptake but did not significantly alter the effect on the frontal cortex. Irradiation with continuous-wave microwaves did not significantly affect choline uptake in the hippocampus, striatum, and hypothalamus but decreased the uptake in the frontal cortex. The effect on the frontal cortex was not altered by pretreatment with narcotic antagonist. These data suggest that exposure to low-level pulsed or continuous-wave microwaves leads to changes in cholinergic functions in the brain.

The changes in heating patterns due to perturbations by thermometer probes in microwave fields were investigated in static phantoms at 915 and 434 MHz. Thermograms taken parallel to the plane of E and H fields, at depths of up to 2 cm, indicated heating changes of +25 to -45 per cent at 915 MHz and +/- 15 per cent at 434 MHz. The amount of perturbation is dependent on the length, size and location of the probes in the RF fields and their orientations to the electric field. If proper probe placement techniques are not observed when metallic probes are involved, hot and cool spots can be generated and shifted to sites that are not measured. Therefore misleading temperatures can result when changes in heating patterns are not detected. Perturbation also varies with applicator designs and phantom geometry. If thermistors and thermocouples are used, the effects of perturbation should be investigated with individual applicators under applicable clinical conditions.