High intensity focused ultrasound combined with ultrasound-guided suction curettage treatment for cesarean scar pregnancy: a comparison of different HIFU sonication strategies.

OBJECTIVE: To explore a new high-intensity focused ultrasound (HIFU) sonication strategy for cesarean scar pregnancy (CSP) and to compare the clinical effectiveness and safety of this new HIFU sonication strategy with the conventional HIFU sonication strategy followed by ultrasound-guided dilation and curettage (USg-D&C) for CSP. MATERIALS AND METHODS: 91 patients with CSP treated by HIFU and USg-D&C in People's Hospital of Deyang City between January 2017 and December 2019 were retrospectively reviewed in this study. Based on the HIFU sonication strategy, patients were divided to two groups: 44 patients were exposed to 'C-shape' sonication layer by layer around the implantation location of the pregnancy sac (control group), while the other 47 patients were exposed to 'I-shape' sonication layer by layer only on the deep part which close to the bladder of the implantation location of the pregnancy sac (experimental group). The differences in clinical efficacy between the two groups were analyzed. Baseline characteristics, technical parameters of HIFU treatment and USg-D&C data were recorded. Adverse events were also recorded. RESULTS: No statistically significant difference was observed between the two groups in baseline characteristics including age, body mass index (BMI), menopause time, largest diameter of gestational sac, pretreatment serum ß-hCG, thickness of gestational sac, embedding myometrium, previous cesarean sections and interval from last cesarean section (CS). The average treatment intensity in the experimental group was significantly lower than that in the control group (p < .05). The median sonication time, total energy used for HIFU ablation, and energy efficiency factor (EEF) in the experimental group were significantly lower than the control group (p < .05). No statistically significant difference was observed between the two groups in treatment power and treatment time (p > .05). Sciatic/buttock pain and postoperative lower abdominal pain in the control group were significantly stronger than that in the experimental group (p < .05). There were no statistically significant differences in post-HIFU vaginal bleeding and discharging, urinary tract irritation, the operation time of USg-D&C, the amount of vaginal bleeding during USg-D&C, and the time for serum ß-hCG back to a normal level between the two groups (p > .05). CONCLUSIONS: The 'I-shape' strategy of HIFU treatment for CSP was effective and safe, with shorter sonication time, less energy input and lower incidence of sonication-related pain occurred in postoperative lower abdominal and sciatic nerve/buttock.

Evaluation of efficacy of new lens nucleus fragmenting forceps in cataract surgery / Avaliação da eficácia de uma nova pinça fragmentadora de núcleo do cristalino para cirurgia de catarata

ABSTRACT Objective: To evaluate the efficacy of low-cost nucleus fragmenting forceps designed to reduce the use of ultrasound during phacoemulsification. Methods: A total of 60 pig eyes enucleated 10 hours before were placed in the microwave oven, at maximum power for 10 seconds, to form cataracts with hardness comparable to a grade IV nucleus in the Lens Opacities Classification System III. Cataract extraction was performed using the Centurion® phacoemulsifier (Alcon Laboratories, Geneve, Switzerland) and Leica M620 microscope. All eyes were submitted to the pre-chop technique dividing the nucleus into four parts. After the pre-chop, the quadrants in 30 eyes were phacoemulsified with the torsional mode and were fragmented in the remaining 30 eyes after the pre-chop was with the fragmentation forceps before torsional mode phacoemulsification. The device was calibrated for all eyes by applying the following parameters: 40% linear torsional phacoemulsification; intraocular pressure of 65 mmHg; the linear vacuum of 600 mmHg; aspiration flow of 40 ccs/minute. After each procedure, the following was recorded: cumulative dissipated energy; equivalent average torsional amplitude; equivalent average ultrasonic power; estimated aspirated fluid; ultrasound total time; and total aspiration time. Statistical analysis was performed using the Kruskal-Wallis test and the IBM Statistical Package for Social Sciences. The p-value <0.05 was considered statistically significant. Results: There was a statistically significant reduction favoring the use of the nucleus fragmenting forceps in all parameters, except for the average torsional amplitude. Conclusion: The use of the nucleus fragmenting forceps contributed to improving the efficacy of torsional phacoemulsification in enucleated pig eyes.

RESUMO Objetivo: Avaliar a eficácia de uma pinça fragmentadora de núcleo, de baixo custo, desenvolvida para reduzir o uso de ultrassom durante a emulsificação do núcleo. Métodos: Sessenta olhos de porco com 10 horas de enucleação foram colocados no forno de microondas, em potência máxima por 10 segundos, para a formação de catarata com dureza comparável à de um núcleo grau IV na Lens Opacities Classification System III. A extração da catarata foi realizada com o facoemulsificador Centurion® (Alcon Laboratories, Genebra, Suíça) e microscópio Leica M620. Todos os olhos foram submetidos a técnica de pre-chop, dividindo o núcleo em quatro partes. Em 30 olhos, após o pre-chop, foi feita a facoemulsificação dos quadrantes com o modo torsional e, nos outros 30 olhos, após o pre-chop, cada quadrante foi fragmentado com a pinça antes da facoemulsificação com o modo torsional. O aparelho foi calibrado para todos os olhos com os seguintes parâmetros: faco torsional linear 40%; pressão intraocular 65 mmHg; vácuo linear 600mmHg e fluxo de aspiração 40cc/minuto. Após cada procedimento, verificaram-se energia dissipada acumulada; média da amplitude do faco torsional; média equivalente do poder ultrassônico; líquido aspirado estimado; tempo total de ultrassom e tempo total de aspiração. A análise estatística foi realizada utilizando o teste de Kruskal-Wallis com o IBM Statistical Package for Social Sciences. Valor de p<0,05 foi considerado estatisticamente significante. Resultados: Houve redução estatisticamente significante em favor do uso da pinça fragmentadora de núcleo em todos os parâmetros, menos na média de amplitude do faco torsional. Conclusão: O uso da pinça fragmentadora de núcleo contribuiu para melhorar a eficácia do faco torsional em olhos de porco enucleados.

Blood-brain barrier disruption and delivery of irinotecan in a rat model using a clinical transcranial MRI-guided focused ultrasound system.

We investigated controlled blood-brain barrier (BBB) disruption using a low-frequency clinical transcranial MRI-guided focused ultrasound (TcMRgFUS) device and evaluated enhanced delivery of irinotecan chemotherapy to the brain and a rat glioma model. Animals received three weekly sessions of FUS, FUS and 10 mg/kg irinotecan, or irinotecan alone. In each session, four volumetric sonications targeted 36 locations in one hemisphere. With feedback control based on recordings of acoustic emissions, 98% of the sonication targets (1045/1071) reached a pre-defined level of acoustic emission, while the probability of wideband emission (a signature for inertial cavitation) was than 1%. BBB disruption, evaluated by mapping the R1 relaxation rate after administration of an MRI contrast agent, was significantly higher in the sonicated hemisphere (P < 0.01). Histological evaluation found minimal tissue effects. Irinotecan concentrations in the brain were significantly higher (P < 0.001) with BBB disruption, but SN-38 was only detected in <50% of the samples and only with an excessive irinotecan dose. Irinotecan with BBB disruption did not impede tumor growth or increase survival. Overall these results demonstrate safe and controlled BBB disruption with a low-frequency clinical TcMRgFUS device. While irinotecan delivery to the brain was not neurotoxic, it did not improve outcomes in the F98 glioma model.

Focused ultrasound as a novel strategy for noninvasive gene delivery to retinal Müller glia.

Müller glia are specialized retinal cells with stem cell properties in fish and frogs but not in mammals. Current efforts to develop gene therapies to activate mammalian Müller glia for retinal repair will require safe and effective delivery strategies for recombinant adeno-associated viruses (AAVs), vectors of choice for clinical translation. Intravitreal and subretinal injections are currently used for AAV gene delivery in the eye, but less invasive methods efficiently targeting Müller glia have yet to be developed. Methods: As gene delivery strategies have been more extensively studied in the brain, to validate our vectors, we initially compared the glial tropism of AAV-PHP.eB, an AAV9 that crosses the blood-brain and blood-retinal barriers, for its ability to drive fluorescent protein expression in glial cells in both the brain and retina. We then tested the glial transduction of AAV2/8-GFAP-mCherry, a virus that does not cross blood-brain and blood-retinal barriers, for its effectiveness in transducing Müller glia in murine retinal explants ex vivo. For in vivo assays we used larger rat eyes, performing invasive intravitreal injections, and non-invasive intravenous delivery using focused ultrasound (FUS) (pressure amplitude: 0.360 - 0.84 MPa) and microbubbles (Definity, 0.2 ml/kg). Results: We showed that AAV-PHP.eB carrying a ubiquitous promoter (CAG) and green fluorescent protein (GFP) reporter, readily crossed the blood-brain and blood-retinal barriers after intravenous delivery in mice. However, murine Müller glia did not express GFP, suggesting that they were not transduced by AAV-PHP.eB. We thus tested an AAV2/8 variant, which was selected based on its safety record in multiple clinical trials, adding a glial fibrillary acidic protein (GFAP) promoter and mCherry (red fluorescent protein) reporter. We confirmed the glial specificity of AAV2/8-GFAP-mCherry, showing effective expression of mCherry in astrocytes after intracranial injection in the mouse brain, and of Müller glia in murine retinal explants. For in vivo experiments we switched to rats because of their larger size, injecting AAV2/8-GFAP-mCherry intravitreally, an invasive procedure, demonstrating passage across the inner limiting membrane, leading to Müller glia transduction. We then tested an alternative non-invasive delivery approach targeting a different barrier - the inner blood-retinal-barrier, applying focused ultrasound (FUS) to the retina after intravenous injection of AAV2/8 and microbubbles in rats, using magnetic resonance imaging (MRI) for FUS targeting. FUS permeabilized the rat blood-retinal-barrier and allowed the passage of macromolecules to the retina (Evans blue, IgG, IgM), with minimal extravasation of platelets and red blood cells. Intravenous injection of microbubbles and AAV2/8-GFAP-mCherry followed by FUS resulted in mCherry expression in rat Müller glia. However, systemic delivery of AAV2/8 also had off-target effects, transducing several murine peripheral organs, particularly the liver. Conclusions: Retinal permeabilisation via FUS in the presence of microbubbles is effective for delivering AAV2/8 across the inner blood-retinal-barrier, targeting Müller glia, which is less invasive than intravitreal injections that bypass the inner limiting membrane. However, implementing FUS in the clinic will require a comprehensive consideration of any off-target tropism of the AAV in peripheral organs, combined ideally, with the development of Müller glia-specific promoters.

Avaliação do método diagnóstico vórtex e sonicação nas infecções de corrente sanguínea relacionadas à cateteres em pacientes portadores do vírus da imunodeficiência humana / Evaluation of the method vortex and sonication in the diagnosis of central line associated bloodstream infection among people with human immunodeficiency virus

Introdução: O uso do cateter venoso central (CVC) é comum no ambiente hospitalar, porém está associado à formação de biofilmes, levando a infecção primária de corrente sanguínea associada à cateter (IPCS). Os métodos diagnósticos atualmente recomendados para infecção primária de corrente sanguínea relacionada a cateter (ICSRC) são rolamento, sonicação e hemocultura. A sensibilidade e especificidade de tais métodos foram testados na população geral, entretanto não sabemos qual o comportamento na população vivendo com vírus da imunodeficiência humana (PVHIV). De tal forma, avaliamos os métodos vórtex -sonicação-vórtex e o comparamos com rolamento e hemocultura. Objetivo: comparar acurácia dos métodos rolamento do cateter, vórtex-sonicação-vórtex e hemoculturas em frascos, para o diagnóstico de infecção de corrente sanguínea relacionado à cateter (ICSRC), na PVHIV adulta no Instituto de Infectologia Emilio Ribas (IIER). Metodologia: Estudo piloto para comparação da acurácia de métodos diagnósticos de ICSRC na PVHIV nos meses de outubro e novembro de 2019, de acordo com os critérios da ANVISA, 2017. Resultados: Avaliamos 10 cateteres pertencentes a 9 pacientes. Das 10 amostras foram isolados os seguintes agentes: CoNs, E.coli, E.faecium, E.faecalis, K.pneumoniae, A.lwoffii e A.haemolyticus, porém nem todas preenchiam critério de definição para ICSRC. O diagnóstico pelo método vórtex-sonicação-vórtex, assim como o rolamento, não se fez presente em nenhum dos casos, apesar de microorganismos terem sido isolados por esses métodos, pois não preencheram os critérios preconizados pela ANVISA. Duas amostras (20%) alcançaram o critério para ICSRC e as outras oito (80%) somente suspeita de IPCS. A acurácia do rolamento e hemocultura, quando comparada a sonicação, foi de 0,8 e 1,0 respectivamente. Os agentes responsáveis por ICSRC no nosso trabalho foram: Acinetobacter lwoffii e Acinetobacter haemolyticus. Não foi possível realizar análise de associação devido ao pequeno número de cateteres no estudo piloto. Conclusão: Observamos em nosso estudo piloto que o método vórtex-sonicação-vórtex isolou mais microorganismos, do que o rolamento, porém por nenhum dos dois métodos foi possível preencher o critério de ICSRC na população portadora do vírus da imunodeficiência humana. O critério inclui o isolamento do agente por um desses métodos, respeitando o cut-off de cada um, associado a hemocultura com isolamento do mesmo agente. São necessários mais estudos com número 6 maior de participantes para avaliação da acurácia dos métodos, bem como a definição do cut-off para essa população para melhor sensibilidade e especificidade dos métodos diagnósticos

Unexpected positive cultures in presumed aseptic revision spine surgery using sonication.

AIMS: During revision procedures for aseptic reasons, there remains a suspicion that failure may have been the result of an undetected subclinical infection. However, there is little evidence available in the literature about unexpected positive results in presumed aseptic revision spine surgery. The aims of our study were to estimate the prevalence of unexpected positive culture using sonication and to evaluate clinical characteristics of these patients. PATIENTS AND METHODS: All patients who underwent a revision surgery after instrumented spinal surgery at our institution between July 2014 and August 2016 with spinal implants submitted for sonication were retrospectively analyzed. Only revisions presumed as aseptic are included in the study. During the study period, 204 spinal revisions were performed for diagnoses other than infection. In 38 cases, sonication cultures were not obtained, leaving a study cohort of 166 cases. The mean age of the cohort was 61.5 years (sd 20.4) and there were 104 female patients. RESULTS: Sonication cultures were positive in 75 cases (45.2%). Hardware failure was the most common indication for revision surgery and revealed a positive sonication culture in 26/75 cases (35%) followed by adjacent segment disease (ASD) in 23/75 cases (30%). Cutibacterium acnes and Staphylococcus epidermidis were the most commonly isolated microorganisms, observed in 45% and 31% of cases, respectively. C. acnes was isolated in 65.2% of cases when the indication for revision surgery was ASD. CONCLUSION: Infection must always be considered as a possibility in the setting of spinal revision surgery, especially in the case of hardware failure, regardless of the lack of clinical signs. Sonication should be routinely used to isolate microorganisms adherent to implants. Cite this article: Bone Joint J 2019;101-B:621-624.

High frequency of low-virulent microorganisms detected by sonication of pedicle screws: a potential cause for implant failure.

OBJECTIVE: Loosening of pedicle screws is a frequent complication after spinal surgery. Implant colonization with low-virulent microorganisms forming biofilms may cause implant loosening. However, the clinical evidence of this mechanism is lacking. Here, the authors evaluated the potential role of microbial colonization using sonication in patients with clinical pedicle screw loosening but without signs of infection. METHODS: All consecutive patients undergoing hardware removal between January 2015 and December 2017, including patients with screw loosening but without clinical signs of infection, were evaluated. The removed hardware was investigated using sonication. RESULTS: A total of 82 patients with a mean (± SD) patient age of 65 ± 13 years were eligible for evaluation. Of the 54 patients with screw loosening, 22 patients (40.7%) had a positive sonication result. None of the 28 patients without screw loosening who served as a control cohort showed a positive sonication result (p < 0.01). In total, 24 microorganisms were detected in those 22 patients. The most common isolated microorganisms were coagulase-negative staphylococci (62.5%) and Cutibacterium acnes (formerly known as Propionibacterium acnes) (25%). When comparing only the patients with screw loosening, the duration of the previous spine surgery was significantly longer in patients with a positive microbiological result (288 ± 147 minutes) than in those with a negative result (201 ± 103 minutes) (p = 0.02). CONCLUSIONS: The low-virulent microorganisms frequently detected on pedicle screws by using sonication may be an important cause of implant loosening and failure. Longer surgical duration increases the likelihood of implant colonization with subsequent screw loosening. Sonication is a highly sensitive approach to detect biofilm-producing bacteria, and it needs to be integrated into the clinical routine for optimized treatment strategies.

Time-domain simulation of ultrasound propagation with fractional Laplacians for lossy-medium biological tissues with complicated geometries.

Simulations of ultrasound wave propagation inside biological tissues have a wide range of practical applications. In previous studies, wave propagation equations in lossy biological media are solved either with convolutions, which consume a large amount of memory, or with pseudo-spectral methods, which cannot handle complicated geometries effectively. The approach described in the paper employed a fractional central difference method (FCD), combined with the immersed boundary (IB) method for the finite-difference, time-domain simulation. The FCD method can solve the fractional Laplace terms in Chen and Holm's lossy-medium equations directly in the physical domain without integral transforms. It also works naturally with the IB method, which enables a simple Cartesian-type grid mesh to be used to solve problems with complicated geometries. The numerical results agree very well with the analytical solutions for frequency power-law attenuation lossy media.

Frequency bands for ultrasound, suitable for the consideration of its health effects.

It is proposed that the ultrasound frequency spectrum should be divided into three bands in order to facilitate a more rational assessment of its health effects. Whilst statement of the frequencies at the borders of these bands facilitates their definition, it is recognized that these observables vary continuously with frequency and consequently these border frequencies should not be used to rule out the possibility of a given effect occurring. The lowest band, US(A), lies between 17.8 and 500 kHz. In this band acoustic cavitation and its associated forces form the dominant process resulting in biological effects in liquids and soft tissues, whereas health effects from airborne ultrasound have been reported but are far less researched. In the middle band, US(B), between 500 kHz and 100 MHz, temperature rise in tissues becomes the most important biological effect of exposure. The highest band, US(C), covers frequencies above 100 MHz, for which the radiation force becomes an increasingly important biophysical mechanism. A justification for the selection of 17.8 kHz in preference to any other threshold for the lower frequency limit for ultrasound is given.

Modelling the temperature evolution of bone under high intensity focused ultrasound.

Magnetic resonance-guided high intensity focused ultrasound (MR-HIFU) has been clinically shown to be effective for palliative pain management in patients suffering from skeletal metastasis. The underlying mechanism is supposed to be periosteal denervation caused by ablative temperatures reached through ultrasound heating of the cortex. The challenge is exact temperature control during sonication as MR-based thermometry approaches for bone tissue are currently not available. Thus, in contrast to the MR-HIFU ablation of soft tissue, a thermometry feedback to the HIFU is lacking, and the treatment of bone metastasis is entirely based on temperature information acquired in the soft tissue adjacent to the bone surface. However, heating of the adjacent tissue depends on the exact sonication protocol and requires extensive modelling to estimate the actual temperature of the cortex. Here we develop a computational model to calculate the spatial temperature evolution in bone and the adjacent tissue during sonication. First, a ray-tracing technique is used to compute the heat production in each spatial point serving as a source term for the second part, where the actual temperature is calculated as a function of space and time by solving the Pennes bio-heat equation. Importantly, our model includes shear waves that arise at the bone interface as well as all geometrical considerations of transducer and bone geometry. The model was compared with a theoretical approach based on the far field approximation and an MR-HIFU experiment using a bone phantom. Furthermore, we investigated the contribution of shear waves to the heat production and resulting temperatures in bone. The temperature evolution predicted by our model was in accordance with the far field approximation and agreed well with the experimental data obtained in phantoms. Our model allows the simulation of the HIFU treatments of bone metastasis in patients and can be extended to a planning tool prior to MR-HIFU treatments.

Controllable permeability of blood-brain barrier and reduced brain injury through low-intensity pulsed ultrasound stimulation.

It has been shown that the blood-brain barrier (BBB) can be locally disrupted by focused ultrasound (FUS) in the presence of microbubbles (MB) while sustaining little damage to the brain tissue. Thus, the safety issue associated with FUS-induced BBB disruption (BBBD) needs to be investigated for future clinical applications. This study demonstrated the neuroprotective effects induced by low-intensity pulsed ultrasound (LIPUS) against brain injury in the sonicated brain. Rats subjected to a BBB disruption injury received LIPUS exposure for 5 min after FUS/MB application. Measurements of BBB permeability, brain water content, and histological analysis were then carried out to evaluate the effects of LIPUS. The permeability and time window of FUS-induced BBBD can be effectively modulated with LIPUS. LIPUS also significantly reduced brain edema, neuronal death, and apoptosis in the sonicated brain. Our results show that brain injury in the FUS-induced BBBD model could be ameliorated by LIPUS and that LIPUS may be proposed as a novel treatment modality for controllable release of drugs into the brain.

Evaluation of inertial cavitation activity in tissue through measurement of oxidative stress.

Ultrasound cavitation is an essential mechanism involved in the therapeutic local enhancement of drug delivery by ultrasound for cancer treatment. Inertial cavitation also triggers chemical reactions that generate free radicals and subsequent oxidative stress in the tissue. The aim of this study was to measure the oxidative stress induced by inertial cavitation in ex vivo tissue and to test the association between the exposure conditions and the oxidative stress. A confocal ultrasound setup was used to sonicate and create inertial cavitation in freshly excised adipose pig tissue. The ex vivo tissue samples were then processed to measure the quantity of malondialdehyde (MDA), an end-product of polyunsaturated free fatty acid oxidation. The creation of hydroxyterephthalic acid (HTA) from the reaction of terephthalic acid (TA) with free radicals in water was also quantified in vitro. Samples were sonicated for different durations using various amplitudes for the applied pressure. The results showed a minimum 2-fold increase in the amount of detected MDA in the sonicated tissue samples compared to baseline clearly suggesting the generation of free radicals by inertial cavitation. The method exhibited a moderate dependence of MDA generated upon the duration of exposure (R(2)=057,p<0.0001). The average increase in MDA concentration was approximately 2-fold, 5-fold, 6-fold, and 9-fold for exposure durations per unit of volume of 0.13, 0.17, 0.25, and 0.50s/mm(3), respectively. The results showed no statistically significant dependence on the amplitude of the pressure within the used range. Both pressure amplitude and exposure duration, however, influenced the HTA concentration (R(2)>0.95,p<0.0001). This biochemical method can be used on ex vivo tissue to detect the generation of free radicals induced by inertial cavitation. In large enough sample populations, the cavitation activity is linked to the exposure conditions of the sonication.

A pilot study of endoluminal US for stool liquefaction.

BACKGROUND: There is a need to cleanse patients who are poorly prepared for colonoscopy safely and efficiently during the procedure to minimize rescheduling. US is already being used in catheter-based intravascular thrombolysis, and time-reversal acoustic (TRA) has been explored in assisting drug delivery to the brain. OBJECTIVE: To explore the efficacy and safety of a miniaturized endoluminal US device in stool dissolution as a means to salvage poor bowel preparation. DESIGN: Proof of concept experimental study. SETTINGS: Animal laboratory. INTERVENTIONS: Low-frequency US and TRAs. MAIN OUTCOME MEASUREMENTS: Feasibility, efficacy, and safety of US to liquefy stools ex vivo. RESULTS: Depending on parameters, such as pulse rate, acoustic intensity, and duration, increases in liquefaction speeds by a factor of 50 and 100 times were obtained. There was a significant difference in weight change between the 20-kHz-treated sample compared with controls (P ≤ .0001). There was no difference in sloughing of mucosa and mechanical injury among the US, water spray, and control groups. LIMITATIONS: Animal model. CONCLUSION: Endoluminal US can liquefy stools at acoustic exposure levels that do not damage ex vivo colonic mucosa. Endoluminal US should be able to dissolve stools more rapidly than water spray alone, thereby optimizing colonoscopic evaluation in vivo.

Reduction of peak acoustic pressure and shaping of heated region by use of multifoci sonications in MR-guided high-intensity focused ultrasound mediated mild hyperthermia.

PURPOSE: Ablative hyperthermia (>55 °C) has been used as a definitive treatment for accessible solid tumors not amenable to surgery, whereas mild hyperthermia (40-45 °C) has been shown effective as an adjuvant for both radiotherapy and chemotherapy. An optimal mild hyperthermia treatment is spatially accurate, with precise and homogeneous heating limited to the target region while also limiting the likelihood of unwanted thermal or mechanical bioeffects (tissue damage, vascular shutoff). Magnetic resonance imaging-guided high-intensity focused ultrasound (MR-HIFU) can noninvasively heat solid tumors under image-guidance. In a mild hyperthermia setting, a sonication approach utilizing multiple concurrent foci may provide the benefit of reducing acoustic pressure in the focal region (leading to reduced or no mechanical effects), while providing better control over the heating. The objective of this study was to design, implement, and characterize a multifoci sonication approach in combination with a mild hyperthermia heating algorithm, and compare it to the more conventional method of electronically sweeping a single focus. METHODS: Simulations (acoustic and thermal) and measurements (acoustic, with needle hydrophone) were performed. In addition, heating performance of multifoci and single focus sonications was compared using a clinical MR-HIFU platform in a phantom (target = 4-16 mm), in normal rabbit thigh muscle (target = 8 mm), and in a Vx2 tumor (target = 8 mm). A binary control algorithm was used for real-time mild hyperthermia feedback control (target range = 40.5-41 °C). Data were analyzed for peak acoustic pressure and intensity, heating energy efficiency, temperature accuracy (mean), homogeneity of heating (standard deviation [SD], T10 and T90), diameter and length of the heated region, and thermal dose (CEM(43)). RESULTS: Compared to the single focus approach, multifoci sonications showed significantly lower (67% reduction) peak acoustic pressures in simulations and hydrophone measurements. In a rabbit Vx2 tumor, both single focus and multifoci heating approaches were accurate (mean = 40.82±0.12 °C [single] and 40.70±0.09 °C [multi]) and precise (standard deviation = 0.65±0.05 °C [single] and 0.64±0.04 °C [multi]), producing homogeneous heating (T(10-90) = 1.62 °C [single] and 1.41 °C [multi]). Heated regions were significantly shorter in the beam path direction (35% reduction, p < 0.05, Tukey) for multifoci sonications, i.e., resulting in an aspect ratio closer to one. Energy efficiency was lower for the multifoci approach. Similar results were achieved in phantom and rabbit muscle heating experiments. CONCLUSIONS: A multifoci sonication approach was combined with a mild hyperthermia heating algorithm, and implemented on a clinical MR-HIFU platform. This approach resulted in accurate and precise heating within the targeted region with significantly lower acoustic pressures and spatially more confined heating in the beam path direction compared to the single focus sonication method.The reduction in acoustic pressure and improvement in spatial control suggest that multifoci heating is a useful tool in mild hyperthermia applications for clinical oncology.

Pre-treatment of sperm reduces success of ICSI in the pig.

In pigs, although ICSI is a feasible fertilization technique, its efficiency is low. In general, injected pig sperm are insufficient to induce oocyte activation and embryonic development. Pretreatments for disrupting sperm membranes have been applied to improve the fertility of ICSI oocytes; however, we hypothesize that such pretreatment(s) may reduce the ability of the sperm to induce oocyte activation. We first evaluated the effects of sperm pretreatments (sonication (SO) to isolate the sperm heads from the tails, Triton X-100 (TX), and three cycles of repeated freezing/thawing (3×-FT) for disrupting sperm membranes) on the rate of pronucleus (PN) formation after ICSI. We found that oocytes injected with control (whole) sperm had higher rates of PN formation than those obtained after subjecting the sperm to SO, TX, and 3×-FT. The amounts of phospholipase Cζ (PLCζ), which is thought to be the oocyte-activating factor in mammalian sperm, in sperm treated by each method was significantly lower than that in whole untreated sperm. Furthermore, using immunofluorescence, it was found that in pig sperm, PLCζ was localized to both the post-acrosomal region and the tail area. Thus we demonstrated for the first time that sperm pretreatment leads to a reduction of oocyte-activating capacity. Our data also show that in addition to its expected localization to the sperm head, PLCζ is also localized in the tail of pig sperm, thus raising the possibility that injection of whole sperm may be required to attain successful activation in pigs.

Ultrasound energy in phacoemulsification: a comparative analysis of phaco-chop and stop-and-chop techniques according to the degree of nuclear density.

BACKGROUND AND OBJECTIVE: To evaluate the amount of ultrasound energy used, corneal endothelial cell loss, and central corneal thickness using the phaco-chop and stop-and-chop techniques for cataracts with different degrees of nuclear density. PATIENTS AND METHODS: One hundred two eyes of 51 patients with bilateral senile cataract were included. Each eye was randomly assigned to have either phaco-chop or stop-and-chop nucleofractis during phacoemulsification. The groups were divided into two subgroups according to the nuclear density. The effective phacoemulsification time, endothelial cell density, and central corneal thickness were analyzed. RESULTS: The mean effective phacoemulsification time was similar between the groups in moderately dense nuclei (2.17 +/- 1.33 vs 1.33 +/- 1.05 seconds; P = .41). However, the phaco-chop technique required less effective phacoemulsification time than the stop-and-chop technique in dense nuclei (3.86 +/- 4.18 vs 6.70 +/- 5.43 seconds; P = .01). The endothelial cell loss and the central corneal thickness did not vary significantly between the groups. CONCLUSION: The phaco-chop technique requires lower ultrasound energy for nuclear management than the stop-and-chop technique in dense cataracts and the resulting endothelial loss was similar in both techniques.

Potential for occupational exposure to engineered carbon-based nanomaterials in environmental laboratory studies.

BACKGROUND: The potential exists for laboratory personnel to be exposed to engineered carbon-based nanomaterials (CNMs) in studies aimed at producing conditions similar to those found in natural surface waters [e.g., presence of natural organic matter (NOM)]. OBJECTIVE: The goal of this preliminary investigation was to assess the release of CNMs into the laboratory atmosphere during handling and sonication into environmentally relevant matrices. METHODS: We measured fullerenes (C60), underivatized multiwalled carbon nanotubes (raw MWCNT), hydroxylated MWCNT (MWCNT-OH), and carbon black (CB) in air as the nanomaterials were weighed, transferred to beakers filled with reconstituted freshwater, and sonicated in deionized water and reconstituted freshwater with and without NOM. Airborne nanomaterials emitted during processing were quantified using two hand-held particle counters that measure total particle number concentration per volume of air within the nanometer range (10-1,000 nm) and six specific size ranges (300-10,000 nm). Particle size and morphology were determined by transmission electron microscopy of air sample filters. DISCUSSION: After correcting for background particle number concentrations, it was evident that increases in airborne particle number concentrations occurred for each nanomaterial except CB during weighing, with airborne particle number concentrations inversely related to particle size. Sonicating nanomaterial-spiked water resulted in increased airborne nanomaterials, most notably for MWCNT-OH in water with NOM and for CB. CONCLUSION: Engineered nanomaterials can become airborne when mixed in solution by sonication, especially when nanomaterials are functionalized or in water containing NOM. This finding indicates that laboratory workers may be at increased risk of exposure to engineered nanomaterials.

Osmosonication of blackberry juice: impact on selected pathogens, spoilage microorganisms, and main quality parameters.

UNLABELLED: Osmosonication combines ultrasound with nonthermal concentration. It was applied on tropical highland blackberry (Rubus adenotrichus) juice over different periods of time to assess reductions in microorganism and the impact on main quality parameters. This juice had been inoculated with Salmonella spp., Shigella sp., a lactic acid bacterium, yeasts, and molds. It was then sonicated for 5.9 to 34.1 min at 20 kHz and 0.83 W/mL. Nonthermal concentration was simulated by mixing the juice with a concentrate to obtain 650 g TSS/kg. It was then stored at -18 °C for up to 82 h. The lactic acid bacterium, yeasts, and molds were reduced by 1.60 to as much as 5.01 log(10) CFU/mL, whereas, for pathogens, reductions were total ≥7.1 log(10) CFU/mL after 24 h of storage, even for juice not sonicated, because of low pH. Color, antioxidant capacity, anthocyanins, and ellagitannins did not change significantly during sonication treatment up to 32 min. However, an off-flavor was detected after 8 min of sonication. Nonetheless, osmosonication can be considered as an alternative to thermal processes for producing safe and high-quality concentrates. PRACTICAL APPLICATION: Osmosonication represents a potential processing alternative for producing safe and high-quality concentrated fruit juice without applying thermal treatments. Findings reported in this article can also be applied by industries when concentrating juices by classical means at relatively low temperature. It provides industries with a mathematical model specific for blackberry juice, from which different combinations of sonication time and storage time of concentrate can be chosen to achieve safety and quality goals.

Cardiac arrhythmia and injury induced in rats by burst and pulsed mode ultrasound with a gas body contrast agent.

OBJECTIVE: Premature complexes (PCs) in the electrocardiogram (ECG) signal have been reported for myocardial contrast echocardiography and also for burst mode (physical therapy) ultrasound with gas body contrast agents at lower peak rarefactional pressure amplitudes (PRPAs). For contrast echocardiography, irreversibly injured cardiomyocytes have been associated with the arrhythmia. The objective was to determine whether cardiomyocyte injury is associated with the PCs induced by the burst mode at lower PRPAs. METHODS: Anesthetized rats were exposed to focused 1.5-MHz ultrasound in a water bath. Evans blue dye was injected intraperitoneally to stain injured cardiomyocytes, and a perflutren lipid microsphere ultrasound contrast agent was infused intravenously. The continuous burst mode simulated physical therapy ultrasound. Intermittent 2-millisecond bursts, or envelopes of pulses simulating diagnostic ultrasound, were triggered 1:4 at end systole. Premature complexes were observed on ECG recordings, and stained cardiomyocytes were counted in frozen sections. RESULTS: The continuous burst mode produced variable PCs and stained cells above a 0.3-MPa PRPA. The triggered bursts above 0.3 MPa and pulse envelopes above 1.2 MPa produced statistically significant (P < .01) PCs and stained cardiomyocytes. CONCLUSIONS: Irreversible cardiomyocyte injury was associated with the development of PCs for the burst mode and occurred at substantially lower PRPAs than for pulsed ultrasound.

Rapporteur report: Roundup, discussion and recommendations.

There is a relative paucity of recent information regarding the long-term health effects associated with exposure to ultrasound, and to infrasound and low-frequency noise (LFN). For ultrasound, further epidemiological studies are recommended, and priority should be given to studies investigating the effects of handedness and to studies assessing possible subtle effects on brain function. These studies should reflect contemporary practises in diagnostic ultrasound and have sufficiently long follow-up periods to examine the possibility of effects into late adolescence or beyond. In the absence of a non-exposed control group, it would be advisable to make comparisons between a highly exposed group with a less exposed group, and to compare groups exposed at differing gestational stages. The effects associated with ultrasound contrast agents should also be studied, and the appropriateness of the thermal index (TI) and mechanical index (MI) should be reviewed. It is recommended that animal models should be used to investigate the effects of exposure at differing gestational ages on development, and modern cellular and molecular techniques used to investigate potential mechanisms of interaction. Although explicit morphological changes have been reported following occupational and experimental exposures to infrasound and low LFN, it was recommended that a thorough review of the relevant biological and health effects literature was necessary before specific proposals could be made. Uncertainties about the characterisation of these low frequencies also indicated the need to develop appropriate measurement techniques and protocols.