Synergistic bactericidal effects and mechanisms of low intensity ultrasound and antibiotics against bacteria: a review

Low intensity ultrasonic therapy is always an important research area of ultrasonic medicine. This review concentrates on low intensity ultrasound enhancing bactericidal action of antibiotics against bacteria in vitro and in vivo, including planktonic bacteria, bacterial biofilms, Chlamydia, and bacteria in implants. These literatures show that low intensity ultrasound alone is not effective in killing bacteria, while the combination of low intensity ultrasound and antibiotics is promising. Low intensity ultrasound facilitating antibiotic treatment is still in its infancy, and still requires a great deal of research in order to develop the technology on medical treatment scale.     

Using low intensity ultrasound to improve the efficiency of biological phosphorus removal

Low intensity ultrasound can produce various effects on biological materials, such as stimulating enzyme activity, cell growth, biosynthesis, etc., which may improve the efficiency of enhanced biological phosphorus removal (EBPR). We adopt total phosphorus (TP) and dehydrogenase activity (DHA) as indicators to confirm the feasibility of applying low intensity ultrasound in EBPR. Single-factor experiments and orthogonal test were conducted in batch anaerobic/oxic (A/O) process simulation to study the influence of ultrasonic intensity and exposure time in the EBPR process. The results showed that the optimal ultrasonic parameters were 0.2 W/cm² and 10 min under which condition the TP concentration in the effluent was 35–50% lower than that of the control (without ultrasonic irradiation). Changes of sludge activities after ultrasonic irradiation were examined. The improvement of sludge activity by ultrasound took 4 h after irradiation to reach the peak level, when an increase above 50% of DHA has been achieved by ultrasonic irradiation, and the enhancing effects induced by ultrasound disappeared in 16 h after irradiation. A tentative mechanism of biological phosphorus removal enhancement stimulated by ultrasound was discussed based on these phenomena.     

Low intensity pulsed ultrasound for fracture healing: a review of the clinical evidence and the associated biological mechanism of action

Low intensity pulsed ultrasound is used in the clinical treatment of fractures and other osseous defects. Level I clinical studies demonstrate the ability of a specific ultrasound signal (1.5 MHz ultrasound pulsed at 1 kHz, 20% duty cycle, 30 mW/cm² intensity (SATA)) to accelerate the healing time in fresh tibia, radius and scaphoid fractures by up to 40%. Additionally, the same ultrasound signal has been shown to be effective at resolving all types of nonunions of all ages, following a wide range of fracture types and primary fracture management techniques.Recently, significant efforts have resulted in a more comprehensive understanding of the biological mechanism of action that produces the documented clinical outcomes. Low intensity pulsed ultrasound has been demonstrated to accelerate in vivo all stages of the fracture repair process (inflammation, soft callus formation, hard callus formation). In particular, accelerated mineralisation has been demonstrated in vitro with increases in osteocalcin, alkaline phosphatase, VEGF and MMP-13 expression. Integrins, a family of mechanoreceptors present on a wide range of cells involved in the fracture healing process, have been shown to be activated by the ultrasound signal. Downstream of the integrin activation, focal adhesions occur on the surface of cells with the activation of multiple signalling pathways, including the ERK, NF-κβ, and PI3 kinase pathways. These pathways have been directly linked to the production of COX-2 and prostaglandin, which are key to the processes of mineralisation and endochondral ossification in fracture healing.     

Inactivation of microorganisms by low-frequency high-power ultrasound: 1. Effect of growth phase and capsule properties of the bacteria

The aim of this study was to determine the effects of high-intensity low-frequency (20 kHz) ultrasound treatment on the viability of bacteria suspension. More specifically, we have investigated the relationship between the deactivation efficiency and the physical (size, hydrophobicity) and biological (gram-status, growth phase) properties of the microbes. Enterobacter aerogenes, Bacillus subtilis, Staphylococcus epidermidis, S. epidermidis SK and Staphylococcus pseudintermedius were chosen for this study owing to their varying physical and biological properties. The survival ratio of the bacteria suspension was measured as a function of the ultrasound power (up to 13 W) for a constant sonication time of 20 min. Transmission electron microscopy was used to evaluate the ultrasound-induced damages to the microbes. Ultrasound treatment resulted in lethal damage to E. aerogenes and B. subtilis (up to 4.5-log reduction), whereas Staphylococcus spp. were not affected noticeably. Further, E. aerogenes suspensions were more sensitive to ultrasonication in exponential growth phase than when they were in stationary phase. The results of this study demonstrate that the main reason for bacterial resistance to ultrasonic deactivation is due to the properties of the bacterial capsule. Microbes with a thicker and “soft” capsule are highly resistant to ultrasonic deactivation process.     

Modulation of luciferase activity using high intensity focused ultrasound in combination with bioluminescence imaging, magnetic resonance imaging and histological analysis in muscle tissue

This study investigates the effect of high intensity focused ultrasound (HIFU) to muscle tissue transfected with a luciferase reporter gene under the control of a CMV-promoter. HIFU was applied to the transfected muscle tissue using a dual HIFU system. In a first group four different intensities (802 W/cm², 1401 W/cm², 2117 W/cm², 3067 W/cm²) of continuous HIFU were applied 20 s every other week for four times. In a second group two different intensities (802 W/cm², 1401 W/cm²) were applied 20 s every fourth day for 20 times. The luciferase activity was determined by bioluminescence imaging. The effect of HIFU to the muscle tissue was assessed by T1-weighted ± Gd-DTPA, T2-weighted and a diffusion-weighted STEAM sequence obtained on a 1.5-T GE-MRI scanner. Histology of the treated tissue was done at the end. In the first group the photon emission was at 3067.6 W/cm² 1.28 × 10⁷ ± 3.1 × 10⁶ photon/s (5.5 ± 1.2-fold), of 2157.9 W/cm² 8.1 ± 2.7 × 10⁶ photon/s (3.2 ± 1.1-fold), of 1401.9 W/cm² 9.3 ± 1.3 × 10⁶ photon/s (4.9 ± 0.4-fold) and of 802.0 W/cm² 8.6x ± 1.2 × 10⁶ photon/s (4.5 ± 0.6-fold) compared to baseline. In the second group the photon emission was at 1401.9 W/cm² and 802.0 W/cm² 14.1 ± 3.6 × 10⁶ photon/s (6.1 ± 1.5-fold), respectively, 5.1 ± 4.7 × 10⁶ photon/s (6.5 ± 2.0-fold). HIFU can enhance the luciferase activity controlled by a CMV-promoter.     

高强度聚焦超声（HIFU）——一门多学科的研究课题

高强度聚焦超声(high intensity focused ultrasound, 简称HIFU)已作为一种无创外科工具而应用于门诊治疗肿瘤。文章介绍了它的基本原理、有关的研究现状和存在的问题,以及对今后研究工作的建议。文章特别强调高强度聚焦超声是一门多学科的综合研究课题,需要各方面的科学工作者通力协作。     

高强度聚焦超声(HIFU)——一门多学科的研究课题

高强度聚焦超声(high intensity focused ultrasound,简称HIFU)已作为一种无创外科工具而应用于门诊治疗肿瘤。文章介绍了它的基本原理、有关的研究现状和存在的问题,以及对今后研究工作的建议。文章特别强调高强度聚焦超声是一门多学科的综合研究课题,需要各方面的科学工作者通力协作。     

Inactivation of Enterobacter aerogenes in reconstituted skim milk by high- and low-frequency ultrasound

The inactivation of Enterobacter aerogenes in skim milk using low-frequency (20 kHz) and high-frequency (850 kHz) ultrasonication was investigated. It was found that low-frequency acoustic cavitation resulted in lethal damage to E. aerogenes. The bacteria were more sensitive to ultrasound in water than in reconstituted skim milk having different protein concentrations. However, high-frequency ultrasound was not able to inactivate E. aerogenes in milk even when powers as high as 50 W for 60 min were used. This study also showed that high-frequency ultrasonication had no influence on the viscosity and particle size of skim milk, whereas low-frequency ultrasonication resulted in the decrease in viscosity and particle size of milk. The decrease in particle size is believed to be due to the breakup of the fat globules, and possibly to the cleavage of the κ-casein present at the surface of the casein micelles. Whey proteins were also found to be slightly affected by low-frequency ultrasound, with the amounts of α-lactalbumin and β-lactoglobulin slightly decreasing.     

Inactivation of microorganisms by low-frequency high-power ultrasound: 2. A simple model for the inactivation mechanism

A simple theoretical model based on shear forces generated by the collapse of the ultrasound cavities near the surface of a microorganism is proposed. This model requires two parameters which take into account the number of acoustic cavitation bubbles, and the resistance of the cell wall of the microorganism to the shear forces generated by bubble collapse. To validate the model, high-power low frequency (20 kHz) ultrasound was used to inactivate two microorganisms with very different sizes, viz., a bacterium, Enterobacter aerogenes and a yeast, Aureobasidium pullulans. The inactivation ratio was experimentally measured as a function of sonication time for different ultrasound power and for different initial cell numbers. For both E. aerogenes and A. pullulans the Log of the inactivation ratio decreased linearly with sonication time, and the rate of inactivation increased (D-value decreased) with the increase in sonication power. The rate of inactivation was also found, for both microorganisms, to increase with a decrease in the initial cell number. The fits, obtained using the proposed model, are in very good agreement with the experimental data.     

High intensity ultrasound modified ovalbumin: Structure,interface and gelation properties

Influence of high intensity ultrasound (HIUS) on the structure and properties of ovalbumin (OVA) were investigated. It was found that the subunits and secondary structure of OVA did not change significantly with HIUS treatment from the electrophoretic patterns and circular dichroism (CD) spectrum. The amount of free sulfhydryl groups increased and intrinsic fluorescence spectra analysis indicated changes in the tertiary structure and partial unfold of OVA after sonication increased. Compared with the untreated OVA, HIUS treatment increased the emulsifying activity and foaming ability, and decreased interface tension (oil–water and air–water interface), which due to the increased surface hydrophobicity and decreased the surface net charge in OVA, while the emulsifying and foaming stability had no remarkable differences. The increased particle size may be attributed to formation of protein aggregates. Moreover, the gelation temperatures of HIUS-treated samples were higher than the untreated OVA according to the temperature sweep model rheology, and this effect was consistent with the increased in surface hydrophobicity for ultrasound treated OVA. These changes in functional properties of OVA would promote its application in food industry.     

高强度聚焦超声在医学超声领域中的发展与应用

高强度聚焦超声(high intensity focused ultrasound,简称HIFU)作为一种非侵入性、无毒副作用、具有巨大潜力的治疗肿瘤的手段,近年来已经越来越受到国内外学者的广泛关注。HIFU技术能将超声能量聚焦到人体组织的肿瘤内,产生高温,使肿瘤组织温度迅速上升至65℃以上,而发生不可逆转的凝固性坏死,从而达到消融肿瘤组织的目的。目前中国的HIFU肿瘤治疗技术和临床居国际领先。文章对HIFU在医学超声领域中的发展历史、现状和前景进行了探讨。     

高强度聚焦超声在医学超声领域中的发展与应用

高强度聚焦超声（high intensity focused ultrasound,简称HIFU）作为一种非侵入性、无毒副作用、具有巨大潜力的治疗肿瘤的手段,近年来已经越来越受到国内外学者的广泛关注。HIFU技术能将超声能量聚焦到人体组织的肿瘤内,产生高温,使肿瘤组织温度迅速上升至65℃以上,而发生不可逆转的凝固性坏死,从而达到消融肿瘤组织的目的。目前中国的HIFU肿瘤治疗技术和临床居国际领先。文章对HIFU在医学超声领域中的发展历史、现状和前景进行了探讨。     

Ultrasonic stimulation of the brain to enhance the release of dopamine – A potential novel treatment for Parkinson’s disease

Parkinson’s disease (PD) is characterized by the decrease of dopamine (DA) production and release in the substantia nigra and striatum regions of the brain. Transcranial ultrasound has been exploited recently for neuromodulation of the brain in a number of fields. We have stimulated DA release in PC12 cells using low-intensity continuous ultrasound (0.1 W/cm² − 0.3 W/cm², 1 MHz), 12 h after exposure at 0.2 W/cm², 40 s, the amount of DA content eventually increased 78.5% (p = 0.004). After 10-day ultrasonic treatment (0.3 W/cm², 5 min/d), the DA content in the striatum of PD mice model restored to 81.07% of the control (vs 43.42% in the untreated PD mice model). In addition to this the locomotion activity was restored to the normal level after treatment. We suggest that the low intensity ultrasound-induced DA release can be attributed to a combination of neuron regeneration and improved membrane permeability produced by the mechanical force of ultrasound. Our study indicates that the application of transcranial ultrasound applied below FDA limits, could provide a candidate for relatively safe and noninvasive PD therapy through an amplification of DA levels and the stimulation of dopaminergic neuron regeneration without contrast agents.     

酶电极上葡萄糖氧化酶的活性的X射线微区分析

利用X射线微区分析, 对二氧化硅溶胶-凝胶包埋于普鲁士蓝修饰玻碳电极上的葡萄糖氧化酶的活性进行了分析; 以Ce(NO₃)₃为捕捉剂, 底物葡萄糖经葡萄糖氧化酶作用产生过氧化氢,后者与捕捉剂反应生成沉淀于酶的活性部位。从X射线微区分析结果表明： 酶电极表面固定化酶的分布均匀,且保存较高的酶活,从微观的角度说明了酶电极的性能与酶电极表面酶活分布的关系。此法制备的葡萄糖氧化酶电极具有较高的灵敏度,稳定性,这与电化学测试结果是一致的。     

相控阵高强度聚焦超声的研究进展

相控阵高强度聚焦超声（high Intensity focused ultrasound,HIFU）技术可以通过电子调相自由控制聚焦区域中焦点的形状、位置、个数等,实现高精度、高效率的治疗。文章主要介绍了相控聚焦超声的原理、阵型设计、声场优化、控制算法、电路设计以及换能器材料等几个方面。     

Progress in stacked piezocomposite ultrasonic transducers for low frequency applications

A stacked ultrasonic transducer comprises multiple individual layers connected mechanically in series and electrically in parallel to reduce the fundamental thickness mode resonance to a frequency corresponding to the transit time of the complete stack and the electrical impedance to a value which corresponds to that of the layers of the stack in parallel. In turn, this allows lower frequency resonant operation than would be possible with a single layer, and facilitates electrical impedance matching to typical transmission circuitry. On transmission, an ideal stack of uniform layers will have an output amplitude larger than that of the equivalent single layer by a factor equal to the . However, using conventional signal amplification circuitry on reception, the output voltage amplitude will be smaller than that of the equivalent single layer by a similar factor. In the past, stacks have commonly been assembled from layers of conventional piezoceramic material but more recently there have been reports of stacks of 1–3 piezocomposites and it is this type that is considered here. The work described in this paper is motivated by the need to operate at frequencies lower than are possible using conventional piezocomposite fabrication technology. Progress in comparison of experimental and simulated results is outlined and the highlights of a theoretical design study are presented. These show that although the general behaviour of a stacked structure is easily predicted, a rigorous theoretical analysis is essential to understand the detail of even a limited range of possible designs.