Shin-ichiro Umemura

Mechanism of Cell Damage by Ultrasound in Combination with Hematoporphyrin

The mechanism of cell damage by ultrasound in combination with hematoporphyrin was studied. Mouse sarcoma 180 cell suspensions were exposed to ultrasound for up to 60 s in the presence and absence of hematoporphyrin, with and without active oxygen scavengers. The cell damage enhancement by hematoporphyrin was suppressed by adding histidine but not by mannitol. The enhancement was doubled in rate by substitution of deuterium oxide medium for normal water. Sonoluminescence was produced in a saline solution under similar acoustic conditions and observed to have spectral components that can excite hematoporphyrin molecules. These results suggest that cell damage enhancement is probably mediated via singlet oxygen generated by ultrasonically activated hematoporphyrin.

Synergistic Effect of Ultrasound and Hematoporphyrin on Sarcoma 180

The antitumor effects of combined use of ultrasound (US) and a photosensitizer, hematoporphyrin (Hp), were determined in mice bearing sarcoma 180. In order to find the optimum timing of the US irradiation after the administration of Hp, the Hp concentrations in the tumor and in the plasma were determined and were analyzed pharmacokinetically. Antitumor effects were evaluated by measuring the tumor size and the tumor weight. Hp alone showed no antitumor effect but US alone showed a slight antitumor effect. The combined treatment with US and Hp showed marked synergistic effects on sarcoma 180 (inhibition ratio was 74% of the control). From these results, the enhancement of antitumor effect is thought to be caused by the sensitization of tumor cells to US mediated by Hp.

Concentration and Fractionation of Small Particles in Liquid by Ultrasound

The efficacy of the ultrasonic standing plane wave in concentrating small particles was theoretically evaluated and compared with experimental results. Acoustic energy density was estimated by measuring the ultrasonic absorption, and particle distribution was observed by dark-field microscopy. The theory predicts that diffusion is negligible in concentrating polystyrene spheres larger than 5 µ m in diameter when they are subjected to 2 J/m3 ultrasound. The half-width of the steady-state particle distribution was of the same order of magnitude as the theoretical value for the particles of 1 µ m and 2 µ m diameter. We also applied this concentrating technique to fractionation of polystyrene spheres 10 µ m in diameter, and more than 90% of the particles in the laminar flow were successfully collected.

Enhancement of Ultrasonically Induced Cell Damage by a Gallium‐Porphyrin Complex, ATX‐70

Enhancement of ultrasonically induced cell damage by a gallium‐porphyrin complex [ATX‐70, 2,4‐ bis(l‐decyloxyethyl)‐Ga(III)‐1,3,5,8‐tetramethylporphyrin‐6,7‐dipropionyl diaspartic acid] was investigated. The rate of damage to isolated sarcoma 180 cells in air‐saturated suspension induced by 2 MHz ultrasound irradiation was enhanced more than four times by 80 μM ATX‐70 in contrast to only twice by the same concentration of hematoporphyrin (Hp). The enhancement was almost completely inhibited in the presence of 10 mM histidine in the suspension, but not at all by 100 mM mannitol, which suggests that the enhanced cell damage was mostly mediated by singlet oxygen. Ultrasonically induced active oxygen generation in an air‐saturated aqueous solution of ATX‐70 was studied by detecting the electron spin resonance signals of 2,2,6,6,‐tetramethyl‐4‐piperidone‐N‐oxyl produced by the reaction of 2,2,6,6‐tetramethyl‐4‐piperidone with the generated active oxygen species. The rate of ultrasonically induced nitroxide generation was enhanced five times by 80 μM ATX‐70 in contrast to only twice by Hp. The enhancement was inhibited significantly in the presence of 10 mM histidine in the suspension, but not at all by 100 mM mannitol. The singlet oxygen generation in air‐saturated aqueous solution was further confirmed by the bleaching of N, N‐dimethyl‐4‐nitrosoaniline in the presence of imidazole. The ultrasonically induced bleaching rate was enhanced six times by ATX‐70, in contrast to only twice by Hp.

Near-infrared finger vein patterns for personal identification

We have demonstrated a personal identification system that is based on near-infrared finger vein patterns. Finger vein patterns of 678 volunteers are acquired by transmitting near-infrared light through a finger and capturing the image with a CCD camera. These vein patterns are enhanced by a background-reduction filter. The similarity between two patterns is then quantified by use of the normalized maximum of the cross correlation of the two images after correction of the tilt angle. The enhanced finger vein pattern enabled 678 persons to be successfully identified.

In vitro and in vivo enhancement of sonodynamically active cavitation by second-harmonic superimposition

Acoustic cavitation, the primary mechanism of sonochemical effects, is known to be induced more easily by standing waves than by progressive waves. It has been found that acoustic cavitation can be an order of magnitude enhanced by superimposing the second harmonic on the fundamental. Significant synergistic effects between the fundamental and the second harmonic were observed in both in vitro and in vivo experiments employing a progressive wave field. Second-harmonic superimposition induces in vitro sonochemical reaction as well as fractional harmonic emission at a relatively low ultrasonic intensity even in a progressive wave field. The effect of second-harmonic superimposition was also investigated using exteriorized mouse livers suspended in degassed saline. The intensity threshold for the production of focal tissue damage, paired with fractional harmonic emission was significantly lowered by second-harmonic superimposition especially when a sonodynamically active agent had been administered to the mouse. Insonation with second-harmonic superimposition in combination with such administration may have potential use for selective tumor treatment.

The sector-vortex phased array: acoustic field synthesis for hyperthermia

A sector-vortex phased array capable of generating directly annular-shape foci is analyzed. By driving the sectors of the array with signals whose phase rotates M times around the circular track, annular foci with the same acoustic-signal-phase rotation are produced in the geometrical focal plane of the array. Because of this phase modulation around the focal annuli and the resulting high-spatial-frequency content, the produced acoustic fields are free from secondary foci both behind and in front of the focal plane. The diameter of the focal annuli can be increased by increasing the mode number M. By providing the array with multiple tracks, it is possible to get larger focal annuli than with a single track with the same mode number M. It is also possible to achieve some control of the power deposition patterns in the depth direction in this way. Using a dual track sector-vortex array with practical ranges of the aperture size and number of elements, acoustic power deposition patterns capable of heating the peripheral and central regions of a nonsuperficial tumor a few centimeters in diameter are obtained by computer simulation.<<ETX>>

Enhancement of sonodynamic tissue damage production by second-harmonic superimposition: theoretical analysis of its mechanism

Among the nonthermal effects of ultrasound, acoustic cavitation may have the highest potential for therapeutic applications if it can be somehow controlled. Recent in vitro and in vivo experiments have demonstrated that sonochemically active cavitation can be enhanced an order of magnitude by superimposing the second harmonic onto the fundamental in insonation. Moreover, they have shown that sonochemically active cavitation can be controlled with relative ease, thereby even in a progressive wave field. The effect of second-harmonic superimposition on the rectified diffusion through the gas-liquid interface of cavitated microbubbles is estimated theoretically. The theoretical rectified diffusion rate explained an asymmetric behavior of the threshold for producing sonodynamic tissue damage as a function of the fundamental and the second-harmonic amplitudes. The tissue damage was produced with a focused progressive wave in a liver lobe of a mouse administered with a sonodynamically active agent. The result suggests that the acceleration of the rectified diffusion is a primary mechanism of the enhancement of sonodynamically effective cavitation by second-harmonic superimposition.

Sonodynamically induced effect of rose bengal on isolated sarcoma 180 cells

Purpose: The ultrasonically induced effect of rose bengal (RB) on isolated tumor cells was investigated. Methods: Sarcoma 180 cells were suspended in air-saturated phosphate-buffered saline and exposed to ultrasound in standing wave mode for up to 60 s in the presence and absence of RB. Cell viability was determined by the ability to exclude trypan blue. Results: The rate of inducing cell damage by ultrasound was enhanced two to three times with 160 μM RB, while no cell damage was observed with RB alone. This enhancement was significantly inhibited by histidine. Conclusions: Ultrasonically induced in vitro cell damage was significantly enhanced by RB. A sonochemical mechanism may be suggested since the enhancement was significantly inhibited by an active oxygen scavenger.

Sonochemical activation of hematoporphyrin: An ESR study

The production of 2,2,6,6-tetramethyl-4-piperidone-N-oxyl by reaction of 2,2,6,6-tetramethyl-4-piperidone (TMPone) with ultrasonically generated active species in oxygenated solutions of hematoporphyrin (Hp) was studied by electron spin resonance spectroscopy. The nitroxide production rate in air-saturated TMPone solutions in phosphate-buffered saline of pH 9.0 was significantly higher in the presence of Hp than in its absence. The enhancement of nitroxide production by Hp was significantly inhibited in the presence of sodium azide or histidine in the solution. The production rate with Hp was doubled by substitution of deuterium oxide, while the rate without Hp increased only modestly. These results suggest that a substantial amount of active oxygen can be generated by ultrasound in aqueous solutions of Hp. Since the production rate was not reduced by mannitol and no nitroxide was produced in nitrogen-saturated solutions, it appears that hydroxyl radicals do not account for a major portion of the active oxygen species which reacted with TMPone to yield a nitroxide.