Biomedical subjects
Publications and source records attributed to S Z Child.
A recent report suggested that exposures of land snails to 0.1 mT, 60 Hz magnetic fields for periods of 48-120 h increase mortality levels by 2-10 times. In direct experimental tests, we were unable to confirm this effect.
The threshold for killing of freshly hatched Drosophila larvae exposed to continuous-wave (CW) ultrasound shows a minimum at approximately 0.3 MHz. This suggests that the stiffness of the material surrounding the gas bodies in the organism is comparable to water. From this, it is apparent that the gas bodies in three-day-old larvae that we have used in earlier studies are far larger than resonance size at the frequencies (1-5 MHz) used. Yet, these larvae were killed by short exposures to low-temporal-average-intensity pulsed ultrasound with peak intensities of the order of 10 W/cm2. Hence, it appears that "large" bubbles cannot be ignored in considerations of the biological effects of pulsed ultrasound and lithotripsy.
Tests with mice show that lung tissue is not selectively heated in comparison with other soft tissues and that lung has an efficient mechanism for dissipating that heat which is produced. Temperatures measured with a fine thermocouple placed at the outer surface of the mouse lung show approximately 1 degree C total rise in temperature in the living animal for an incident intensity of 1 W/cm2 (4 MHz, unfocused sound field).
To enhance the likelihood of cavitation, pregnant mice were subjected to hyperbaric conditions and quickly returned to atmospheric pressure. Following this treatment, they were exposed to spatial average, pulse average intensities of 100 W/cm2 (2.2-MHz, 20-microseconds pulses with a duty cycle of 1/1000 or a temporal average spatial average intensity of 0.1 W/cm2). Fetal weights, deaths and malformations were scored. No statistically significant effects were observed in the offspring.
Chick embryos at 72 h incubation were subjected to three double shock waves from a Wolf Model 2137.50 Electrohydraulic Lithotripter. The pressure amplitude at the embryo was adjusted by variation of the distance from the source to the embryo. After a total of 120 h of incubation, they were assessed for developmental abnormalities. Early deaths, delayed deaths and malformations were all significantly increased at pressures of 10 MPa with suggestions of possible effects at lower pressure levels.
Motivated by a recent finding that threshold pressures for hemorrhage in mouse lung exposed to the fields of an electrohydraulic lithotripter were less than 2 MPa, we extended the exposures to pulsed ultrasound. Sharply defined thresholds of the order of 1 MPa were found with 10 microseconds length pulses and roughly twice that value for 1 microsecond pulses. The thresholds at 4 MHz are greater than at 1 MHz. The thresholds are comparable for focused and unfocused fields. As would be expected for a cavitation-like phenomenon, temporal average intensity is a very poor predictor of this effect. In the extreme case, lesions were found at temporal average intensities on the order of 1 mW/cm2.
Threshold pressures for hemorrhage in mouse lung exposed to the fields of an electrohydraulic lithotripter appear to be less than 2 MPa with as few as 10 pulses and with severe damage occurring at levels between 5 and 6 MPa. This is very much smaller than the fields required to fragment kidney and gallstones and smaller than the thresholds for damage to kidney tissues. Fetal lung, in contrast, did not show signs of damage at 20 MPa. The lower sensitivity of fetal lung is consistent with a cavitation-related mechanism for lung damage by shock waves. Since the pressures in these exposures are almost entirely positive, it suggests that the value of negative pressures as predictors of the behavior of gas bodies in tissues should be reconsidered.
A recent study has found that the threshold for extravasation in mouse kidney tissues by exposure to a spark-generated shock wave is of the order of 3-5 MPa (peak positive pressure). Since the mode pressure used by commercial pulsed Doppler ultrasound units is approximately 5 MPa, it is essential to determine whether these observations are relevant to diagnostic ultrasound. Hence, a comparable study has been completed using the same pathological endpoints but with exposure to pulsed ultrasound (10 microseconds pulse length) at 1.2 MHz and 3.8 MHz in which peak positive pressures exceeded 10 MPa. At these levels the focal waves are in shock because of the nonlinear properties of the propagating medium. The results of the pulsed ultrasound study were negative. Although this finding is encouraging for the use of diagnostic ultrasound, the two studies eventually must be integrated into a single mechanistic picture before the limits of safety will be known.
Drosophila larvae contain small gas bodies stabilized within their respiratory system. Because these bubbles are inhibited in their capacity to expand by the surrounding tissues, it is probable that they do not respond to acoustic fields in the manner described by classical cavitation theory that assumes a spherical bubble in an infinite fluid. However, just because of this inhibited expansion, they may serve as reasonable models for the gas bodies in mammalian tissues. Approximately one half of a population of Drosophila larvae is killed by exposure to 3 to 10 double lithotripter shocks with a positive pressure of 2-3 MPa. In contrast with the predictions of classical cavitation theory, adding a negative pressure to the exposure has little influence on the killing rate or its threshold pressure. The available evidence suggests that interaction of gas bodies in tissues with pressure fields and the resultant biological effects may be qualitatively different than predicted by classical cavitation theory and that positive rather than negative pressure may be a predictor of these effects.
Resonance lysis of the cells in the leaves of the aquatic plant Elodea originally reported by Miller have thresholds at intensities of a few W/cm2. With pulsed ultrasound, the resonance behavior vanishes and the thresholds are at much higher amplitudes. This is similar to the characteristics of the thresholds for killing of Drosophila larvae by pulsed ultrasound. Both organisms contain small gas bodies within the tissues which may serve as nuclei for a cavitation related phenomenon. The results suggest that the response of these bubbles to continuous wave fields and very short pulses is qualitatively different.
Comparatively simple analysis shows that diagnostic ultrasound devices, in some cases, may approach output levels that can produce significant heating of tissues, particularly if the propagation path includes bone. Experimental tests of these predictions using rodents show that temperature increments of the order of 3 degrees C/W/cm2 are produced in skull bone with sharply focused fields at medically relevant frequencies.
It has been suggested that fetal weight reduction by ultrasound exposure is linearly related to the dose parameter I2t, where I is the intensity and t the exposure time. A direct test of the concept was conducted using CF-1 mice. No effect on fetal weight was found at values of the dose parameter large enough to produce measurable heating in the fetal and maternal tissues.
Ultrasonographic assessment of ovarian follicular maturity was reportedly associated with atypically early ovulation in women; related studies reported reduced litter sizes in rats. To confirm these findings, mice which were midway between ovulatory gonadotropin (LH or human chorionic gonadotropin) stimulation and ovulation, were sham- or ultrasound-treated periovarially for 5 min. Exposure was at a spatial average, temporal maximum intensity of 60 W/cm2. Carrier frequency in the pulse was 2.2 MHz, pulse length was 10 microseconds, and pulse repetition frequency was 200 Hz. Spatial average, temporal average intensity was 0.12 W/cm2. At autopsy, ultrasound- and sham-treated groups responded similarly in proportions ovulating and in mean ova ovulated. Combined experiments had a 97% chance of detecting a significant (greater than 1 h) advance in ovulation time, had it occurred. Thus, our adequately sensitive mammalian ovulatory tests revealed no association of ultrasound with decrease in ovum number or acceleration in ovulation time (as reported in humans).
Our replicate of a study by Takabayashi et al. (1981) Effects of pulse-wave ultrasonic irradiation on mouse embryo. Cho-Onpa Igaku (Supersonic Medicine) 8, 286-288 failed to show any effects of exposure in utero of mice to spatial average, temporal maximum intensities of 60 W/cm2. Fetuses were exposed at 8 days post fertilization and assessed at 18 days for fetal weight, resorptions, premature deaths, and malformations. Carrier frequency in the pulse was 2 MHz. Pulse lengths of 10 microseconds and pulse repetition frequencies of 1000 and 200 Hz yielded spatial average, temporal average intensities of 0.6 and 0.1 W/cm2. Total exposure time was 5 min. Our results provide no basis to conclude that conditions relevant to human fetal monitoring will cause developmental, externally visible anomalies in mice exposed as 8-day fetuses.
In 1978, Pizzarello and co-workers reported that exposure of rat fetuses to diagnostic levels of ultrasound caused marked reduction in fetal weight. Replicates of these experiments have been conducted and, in addition, exposures at 10 times the peak and average intensity were used. No effect on fetal weight, numbers of living fetuses or resorptions could be attributed to ultrasound at either exposure level.
Isolated stage III forespores of Bacillus megaterium ATCC 19213 in aqueous suspensions were nearly as dehydrated as mature spores, as indicated by low dextran-impermeable volumes of ca. 3.0 ml per g (dry weight) of cells compared with values of ca. 2.6 for mature spores and 7.3 for vegetative cells. The forespores lacked dipicolinate, had only minimal levels of calcium, magnesium, manganese, potassium, and sodium, and were more heat sensitive than vegetative cells. The effective homogeneous conductivities and dielectric constants measured over a frequency range of 1 to 200 MHz indicated that the inherent conductivities of the forespores were unusually low, in keeping with their low mineral contents, but that the forespores could be invaded by environmental ions which could penetrate dielectrically effective membranes. Overall, our findings support the view that the dehydration of a forespore during stage III of sporogenesis may be the result of ion movements out of the forespore into the sporangium.
Drosophila larvae are damaged by exposures to low temporal average intensity pulsed ultrasound with peak intensities of 10-20W/cm2 (2 MHz). Eggs of the same organism are affected by exposures to 3W/cm2 c.w. ultrasound. This experiment shows that eggs become sensitive to high peak intensity (50-100 W/cm2) pulsed ultrasound only shortly before hatching. At this age the larvae have formed and have taken air into the respiratory system. This observation supports the postulate that the sites of action of the ultrasound are the small stabilized gas bodies within the organisms.
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