Sally Z. Child

Lung damage from exposure to pulsed ultrasound

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.

Enhancement of Fibrinolysis With 40-kHz Ultrasound

BACKGROUND Ultrasound at frequencies of 0.5 to 1 MHz and intensities of > or =0.5 W/cm2 accelerates enzymatic fibrinolysis in vitro and in some animal models, but unacceptable tissue heating can occur, and limited penetration would restrict application to superficial vessels. Tissue heating is less and penetration better at lower frequencies, but little information is available regarding the effect of lower-frequency ultrasound on enzymatic fibrinolysis. We therefore examined the effect of 40-kHz ultrasound on fibrinolysis, tissue penetration, and heating. METHODS AND RESULTS 125I-fibrin-radiolabeled plasma clots in thin-walled tubes were overlaid with plasma containing tissue plasminogen activator (tPA) and exposed to ultrasound. Enzymatic fibrinolysis was measured as solubilization of radiolabel. Tissue attenuation and heating were examined in samples of porcine rib cage. Fibrinolysis was increased significantly in the presence of 40-kHz ultrasound at 0.25 W/cm2, reaching 39+/-7% and 93+/-11% at 60 minutes and 120 minutes, compared with 13+/-8% and 37+/-4% in the absence of ultrasound (P<0.0001). The acceleration of fibrinolysis increased at higher intensities. Attenuation of the ultrasound field was only 1.7+/-0.5 dB/cm through the intercostal space and 3.4+/-0.9 dB/cm through rib. Temperature increments in rib were <1 C/(W/cm2). CONCLUSIONS These findings indicate that 40-kHz ultrasound significantly accelerates enzymatic fibrinolysis at intensities of > or =0.25 W/cm2 with excellent tissue penetration and minimal heating. Externally applied 40-kHz ultrasound at low intensities is a potentially useful therapeutic adjunct to enzymatic fibrinolysis with sufficient tissue penetration for both peripheral vascular and coronary applications.

Lysis of erythrocytes by exposure to CW ultrasound

The threshold for lysis of erythrocytes suspended at concentrations of 0.5-1% in saline or plasma in rotating cylindrical exposure vessels is approximately spatial peak intensities of 2 W/cm2 at 1 MHz continuous wave (CW). Results of a series of experiments in which cell concentration, viscosity and gas composition of the suspending medium and rotation speed of the exposure vessel were varied combined with observations of sonoluminescence are all consistent with a hypothesis that cells are lysed by inertial (transient) acoustic cavitation. For the proposed mechanism to operate in cell suspensions, it is necessary that bubbles be brought into contact with the cells. Rotation of the chamber recycles bubbles that are driven by radiation forces to the far wall of the chamber in a matter of milliseconds. The physical and chemical properties of the wall of the chamber appear to be important as stabilizing sites for nuclei that serve as seeds for cavitation events.

Hemolysis in vivo from exposure to pulsed ultrasound

Ultrasonically induced hemolysis in vivo when a commercial ultrasound contrast agent, Albunex, was present in the blood. Murine hearts were exposed for 5 min at either 1.15 or 2.35 MHz with a pulse length of 10 microseconds and pulse repetition frequency of 100 Hz. During the exposure period, four boluses of Albunex were injected into a tail vein for a total of approximately 0.1 mL of Albunex. Following exposure, blood was collected by heart puncture and centrifuged, and the plasma was analyzed for hemoglobin concentration. With Albunex present in the blood, the threshold for hemolysis at 1.15 MHz was 3.0 +/- 0.8 MPa (mean +/- SD) peak positive pressure (approximately 1.9 MPa negative pressure, approximately 180 W cm-2 pulse average intensity). For the highest exposure levels (10 MPa peak positive pressure at the surface of the animal), the mean value for hemolysis was approximately 4% at 1.15 MHz and 0.46% at 2.35 MHz, i.e., the threshold at 2.35 MHz is > 10 MPa peak positive pressure. In contrast, hemolysis in control mice receiving saline injections at 10 MPa or sham-exposed (0 MPa) mice receiving Albunex was approximately 0.4%.

Lung damage from exposure to the fields of an electrohydraulic lithotripter

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.

Effects of ultrasound on drosophila: III. Exposure of larvae to low-temporal-average-intensity, pulsed irradation

Abstract The larvae of Drosophila melanogaster were exposed to repetitive, microsecond length pulses of 2 MHz ultrasound. The observed effects included killing of larvae and delayed lethality in the pupal stage. The effects depended strongly upon the temporal peak intensity with killing observed at spatial. temporal peak intensities above 10 W/cm 2 . In fact, the observations suggest that there may be a threshold for these effects at approximately this intensity. Temporal average intensity appears to be a poor predictor for these biological effects. Marked decreases in survival rates were observed at spatial, temporal average intensities as low as 3 mW/cm 2 .

The influence of contrast agents on hemorrhage produced by lithotripter fields

Ultrasonic contrast agents greatly increase the side effects of low-amplitude lithotripter fields in mice. Using a piezoelectric lithotripter, adult mice were exposed to 200 lithotripter pulses with a peak positive pressure amplitude of 2 MPa. During the exposure period, mice were injected with approximately 0.1 mL of the ultrasonic contrast agent Albunex. For comparison, another group of mice experienced the same lithotripter exposures, but were not injected with contrast agent. Following exposures, animals were sacrificed and observed for hemorrhage in various organs and tissues. Mice exposed to the lithotripter field alone had minimal hemorrhage only in the intestine and lung. In comparison, mice injected with Albunex during exposure exhibited extensive hemorrhage in the intestine, kidney, muscle, mesentery, stomach, bladder, seminal vesicle and fat.

THRESHOLDS FOR ULTRASONICALLY INDUCED LUNG HEMORRHAGE IN NEONATAL SWINE

The threshold for generation of lung hemorrhage in adult mice by pulsed ultrasound has been shown to be approximately 1 MPa at the surface of the lung (10-microseconds pulse and a carrier frequency of 2 MHz). This investigation used neonatal swine to determine if the findings for mice can be generalized to other species. After exploratory observations, the inverse sampling method was used in a primary study (22 animals, 88 exposure sites) to determine the threshold for lung hemorrhage in neonatal swine. The primary study was followed by a separate confirmation study (13 animals, 48 exposure sites), testing the conclusions of the first study and comparing damage at subthreshold levels with sham-exposed animals. A separate investigation explored the histological nature of tissue damage at suprathreshold levels. A 2.3-MHz focused transducer (10 microseconds at 100-Hz pulse-repetition frequency) was incremented vertically for a distance of 2 cm over the chest of the subject for a total exposure period of 16 min. Animals were euthanized and lungs were scored by visual inspection for numbers and areas of gross hemorrhages. The threshold level for hemorrhage was approximately 1.5 MPa peak positive pressure in water at the surface of the animal or, at the surface of the lung, 1.1 MPa peak positive pressure, 1 MPa fundamental pressure, 0.9 MPa maximum negative pressure, 25 W cm-2 pulse average intensity or a mechanical index of 0.6. These values are essentially the same as those reported for adult mice.

Tactile perception of ultrasound.

In this investigation, acoustic radiation force was used as a stimulus to determine the threshold for tactile perception in the human finger and upper forearm as a function of frequency and pulse duration. Initially, a small (1.8-cm2) acoustically reflecting disk was affixed to the anatomical exposure site to maximize the delivered radiation force. Exposures were performed using a 2.2-MHz unfocused source modulated to produce square waves at 50, 100, 200, 500, and 1000 Hz. For the finger, maximum tactile sensitivity occurred at 200 Hz with a threshold radiation force of approximately 0.4 mN. For single pulses of 1 to 100 ms at 2.2 MHz, the threshold forces were an order of magnitude greater than for continuous exposure modulated at 200 Hz. Thresholds for pulse durations of 0.1 ms were somewhat greater than for pulses longer than 1 ms. Subsequently, thresholds of tactile perception were determined for direct exposure of the upper forearm (avoiding bone) to single pulses of 2.2-MHz ultrasound. Comparison of perception thresholds with and without a reflecting material over the tissue were consistent with the hypothesis that the tactile sensation experienced when tissue is exposed to ultrasound is its response to the radiation force associated with the transfer of momentum from the sound field to the tissue medium.

Age dependence of ultrasonically induced lung hemorrhage in mice.

Thresholds for ultrasonically induced lung hemorrhage were determined in neonatal mice (24-36 h old), juvenile mice (14 d old) and adult mice (8-10 weeks old) to assess whether or not the threshold for lung hemorrhage is dependent upon age. Ultrasonic exposures were at 1.15 MHz with a pulse length of 10 microseconds, pulse repetition frequency of 100 Hz and a total exposure duration of 3 min. The threshold for lung hemorrhage occurred at a peak positive acoustic pressure of approximately 1 MPa for mice in all three age groups. Although the thresholds were similar for neonatal, juvenile and adult mice, the sizes of the suprathreshold hemorrhages were significantly larger in adult mice than in neonatal or juvenile mice.