Ultrasound Intensity Research Articles

Low-intensity pulsed ultrasound combined with microbubble mediated JNK/c-Jun pathway to reverse multidrug resistance in triple-negative breast cancer

To investigate the effects of low-intensity pulsed ultrasound combined with microbubble (LIPUS-MB) mediated JNK/c-Jun pathway reversal on multidrug resistance in triple-negative breast cancer and the underlying mechanisms. An orthogonal experiment was designed to screen for the optimal parameters of LIPUS-MB in MDA-MB-231/DOX cells. The CCK-8 assay was used to determine the drug resistance of the cells and to measure their proliferation activity and resistance reversal efficiency at the optimal parameters. Hoechst 33,342 staining and Annexin V-FITC/PI staining were employed to detect cell morphological changes and apoptosis, respectively. The MDA-MB-231/DOX models of transplanted tumor were established in BALB/c. The impact of LIPUS-MB on allograft tumor growth was observed in vivo. Immunohistochemistry was employed to investigate the expression of P-gp, ABCG2, and Ki-67 in tumor tissues, while western blot was utilized to assess the protein expression of P-gp, ABCG2, JNK, p-JNK, c-Jun, p-c-Jun, Bcl-2 and Bax in both MDA-MB-231/DOX cells and allograft tumor tissues. The optimal LIPUS-MB parameters for MDA-MB-231/DOX cells are the microbubble concentration of 20%, ultrasound intensity of 1.0 W/cm2, and irradiation time of 60 s. The drug resistance index of MDA-MB-231/DOX cells is 19.17. Following the optimal parameter application, the IC50 value of the cells decreases by 5.71-fold, with a reversal efficiency of 87.03%, and a simultaneous decrease in cell proliferation activity. Compared with other groups, the DOX + LIPUS-MB group displayed the highest incidence of apoptotic nuclear morphology, and the greatest quantity of cellular apoptosis and the most pronounced decrease in the expression levels of P-gp, ABCG2, p-JNK, p-c-Jun, and Bcl-2 proteins within the cells. Conversely, the expression levels of Bax proteins reach the highest levels (all P < 0.05). Furthermore, in vivo subcutaneous tumor transplantation experiments in nude mice revealed that the DOX + LIPUS-MB group exhibited smaller tumor growth rate, volume and the expression of P-gp, ABCG2, and Ki-67 compared to the DOX + LIPUS group, indicating the most pronounced inhibitory effect on tumor growth and it significantly inhibited tumor proliferation, promoted its apoptosis. In conclusion, following parameter optimization, LIPUS-MB was found to reduce the drug resistance of MDA-MB-231/DOX cells. The underlying mechanism may involve the downregulation of P-gp and ABCG2 proteins expression through the modulation of the JNK/c-Jun pathway by LIPUS-MB, thereby inhibiting cell proliferation activity and promoting apoptosis, and enhancing the in vivo anti-tumor effect of DOX, thus reversing multidrug resistance in triple-negative breast cancer.

An Ultrasound‐Activated Supramolecular Modulator Enhancing Autophagy to Prevent Ventricular Arrhythmias Post‐Myocardial Infarction

AbstractVentricular arrhythmias (VAs) triggered by myocardial infarction (MI) are the leading cause of sudden cardiac mortality worldwide. Current therapeutic strategies for managing MI‐induced VAs, such as left stellate ganglion resection and ablation, are suboptimal, highlighting the need to explore safer and more effective intervention strategies. Herein, we rationally designed two supramolecular sonosensitizers RuA and RuB, engineered through acceptor modification to generate moderate reactive oxygen species (ROS) to modulate VAs. Both RuA and RuB demonstrated high ultrasound (US)‐activated ROS production efficiency, with singlet oxygen (1O2) quantum yield (ΦΔ) of 0.70 and 0.88, respectively, surpassing ligand IR1105 and the conventional sonosensitizer ICG (ΦΔ=0.40). In vitro, RuB, at a modest concentration and under US intensity notably boosts pro‐survival autophagy in microglia BV2 cell. To improve in vivo stability and biocompatibility, RuB was further encapsulated into DSPE‐PEG5000 to prepare RuB nanoparticles (RuB NPs). In vivo studies after microinjection of RuB NPs into the paraventricular nucleus (PVN) and subsequent US exposure, demonstrated that RuB NPs‐mediated US modulation effectively suppresses sympathetic nervous activity (SNA) and inflammatory responses, thereby preventing VAs. Importantly, no tissue injury was observed post RuB NPs‐mediated US modulation. This work pioneers the design of long‐wave emission supramolecular sonosensitizers, offering new insights into regulating cardiovascular diseases.

An Ultrasound-Activated Supramolecular Modulator Enhancing Autophagy to Prevent Ventricular Arrhythmias Post-Myocardial Infarction.

Ventricular arrhythmias (VAs) triggered by myocardial infarction (MI) are the leading cause of sudden cardiac mortality worldwide. Current therapeutic strategies for managing MI-induced VAs, such as left stellate ganglion resection and ablation, are suboptimal, highlighting the need to explore safer and more effective intervention strategies. Herein, we rationally designed two supramolecular sonosensitizers RuA and RuB, engineered through acceptor modification to generate moderate reactive oxygen species (ROS) to modulate VAs. Both RuA and RuB demonstrated high ultrasound (US)-activated ROS production efficiency, with singlet oxygen (1O2) quantum yield (ΦΔ) of 0.70 and 0.88, respectively, surpassing ligand IR1105 and the conventional sonosensitizer ICG (ΦΔ=0.40). In vitro, RuB, at a modest concentration and under US intensity notably boosts pro-survival autophagy in microglia BV2 cell. To improve in vivo stability and biocompatibility, RuB was further encapsulated into DSPE-PEG5000 to prepare RuB nanoparticles (RuB NPs). In vivo studies after microinjection of RuB NPs into the paraventricular nucleus (PVN) and subsequent US exposure, demonstrated that RuB NPs-mediated US modulation effectively suppresses sympathetic nervous activity (SNA) and inflammatory responses, thereby preventing VAs. Importantly, no tissue injury was observed post RuB NPs-mediated US modulation. This work pioneers the design of long-wave emission supramolecular sonosensitizers, offering new insights into regulating cardiovascular diseases.

A Bioengineered Cathepsin B-sensitive Gas Vesicle Nanosystem That Responds With Increased Gray-level Intensity of Ultrasound Biomicroscopic Images

ObjectivesThis work aimed to promote the interaction of a modified Gas Vesicle (GV) with Cathepsin B (CTSB) protease and analyzed their backscattered signal by Ultrasound (US). MethodsWe modified the sequence of the gene coding for GvpC to contain a CTSB cleavage and expressed the protein in an Escherichia coli recombinant system. The protein was purified and added to GVs preparations in which the original GvpC was removed (ΔGV), constituting the modified GV (GV*). Western Blot (WB) test was used to compare GVs with GvpC and engineer GvpC at starting (T0) and after 24 hours (T24) reacting with CTSB. A 21 MHz US B-mode and nonlinear contrast-mode (5% total power) imaged US phantoms having samples of GVwt, ΔGV (stripped GV), GV*, and CTSB+GV*. Also, a 21MHz US B-mode imaged US phantoms having a tumor cell line extracellular fraction (TCEF) and the TCEF+GV* sample. A 100% total US power was applied to collapse the GV structure. ResultsIn WB we detected a decrease in engineered GvpC levels 24 h after the incubation of GV* with CTSB, compared to the concentration at T0, suggesting that CTSB cleaved the engineered GvpC. Regions-of-interest over image of phantom cross-sections were determined and the B-mode image mean gray level intensity resulted in a significant (p<0.05) increase comparing CTSB+GV* with PBS (control), GVwt, ΔGV and GV*. Non-linear mode image gray level intensity from CTSB+GV* increased by 11.79, 7.86 and 14.75 dB from samples containing GVwt, ΔGV and GV*, respectively. GV preparations incubated with TCEF and the TCEF+GV* sample showed an increase of 81% in signal compared to TCEF+GVwt. ConclusionsThe increased ultrasound backscattered signal intensity suggests GVs as a potential biosensor for protease activity, possibly aiding the detection of protease-rich tissue regions.

Stability and cavitation of nanobubble: Insights from large-scale atomistic molecular dynamics simulations.

We perform large-scale atomistic simulations of a system containing 12 × 106 atoms, comprising an oxygen gas-filled bubble immersed in water, to understand the stability and cavitation induced by ultrasound. First, we propose a method to construct a bubble/water system. For a given bubble radius, the pressure inside the bubble is estimated using the Young-Laplace equation. Then, this pressure is used as a reference for a constant temperature, constant pressure simulation of an oxygen system, enabling us to extract a sphere of oxygen gas and place it into a cavity within an equilibrated water box. This ensures that the Young-Laplace equation is satisfied and the bubble is stable in water. Second, this stable bubble is used for ultrasound-induced cavitation simulations. We demonstrate that under weak ultrasound excitation, the bubble undergoes stable cavitation, revealing various fluid velocity patterns, including the first-order velocity field and microstreaming. These fluid patterns emerge around the bubble on a nanometer scale within a few nanoseconds, a phenomenon challenging to observe experimentally. With stronger ultrasound intensities, the bubble expands significantly and then collapses violently. The gas core of the collapsed bubble, measuring 3-4nm, exhibits starfish shapes with temperatures around 1500K and pressures around 6000bar. The simulation results are compared with those from Rayleigh-Plesset equation modeling, showing good agreement. Our simulations provide insights into the stability and cavitation of nanosized bubbles.

Ultrasound‐Assisted Extraction of Bioactive Compounds From Sucupira (Pterodon pubescens) Fruit: Chemical Characterization and Antimicrobial Activity

ABSTRACTThe sucupira fruit (Pterodon pubescens) contains a valuable oil characterized by high levels of fatty acids and an abundance of bioactive compounds, such as flavonoids and terpenoids. This study aimed to evaluate the use of ultrasound‐assisted extraction to obtain sucupira fruit oil through a complete 22 factorial rotational design. The effects of ultrasound intensity (%) and ethanol concentration (%) on lipid content, antioxidant capacity, color, and reducing capacity were assessed. The maximum lipid content was 26.9/100 g, which was significantly higher than that obtained by the Soxhlet method (23.4/100 g). The extract obtained at the optimized conditions (90% ultrasound intensity and 80% ethanol concentration) showed antimicrobial activity against Salmonella and Escherichia coli cocktails. Compounds such as β‐caryophyllene, caryophyllene oxide, α‐humulene, α‐copaene, and spathulenol, responsible for biologically relevant activities, were identified in the extract through GC/GC–MS. HPLC–MS/MS identified compounds such as catechin, luteolin, quercetin, taxifolin, among others.

Interstitial dual-mode ultrasound with a 3-mm MR-compatible catheter for image-guided HIFU and directional in-vitro tissue ablations.

Current interstitial techniques of tumor ablation face challenges that ultrasound technologies could meet. The ablation radius and directionality of the ultrasound beam could improve the efficiency and precision. Here, a 9-gauge MR-compatible dual-mode ultrasound catheter prototype was experimentally evaluated for Ultrasound Image-guided High Intensity Focused Ultrasound (USgHIFU) conformal ablations. The prototype consisted of 64 piezocomposite linear array elements and was driven by an open research programmable dual-mode ultrasound platform. After verifying the US-image guidance capabilities of the prototype, the HIFU output performances (dynamic focusing and HIFU intensities) were quantitatively characterized, together with the associated 3D HIFU-induced thermal heating in tissue phantoms (using MR thermometry). Finally, the ability to produce robustly HIFU-induced thermal ablations in in-vitro liver was studied experimentally and compared to numerical modeling. Investigations of several HIFU dynamic focusing allowed overcoming the challenges of miniaturizing the device: mono-focal focusing maximized deep energy deposition, while multi-focal strategies eliminated grating lobes. The linear-array design of the prototype made it possible to produce interstitial ultrasound images of tissue and tumor mimics in situ. Multi-focal pressure fields were generated without grating lobes and transducer surface intensities reached up to Isapa = 14 W·cm-2. Seventeen elementary thermal ablations were performed in vitro. Rotation of the catheter proved the directionality of ablation, sparing non-targeted tissue. This experimental proof of concept demonstrates the feasibility of treating volumes comparable to those of primary solid tumors with a miniaturized USgHIFU catheter whose dimensions are close to those of tools traditionally used in interventional radiology, while offering new functionalities.

Injectable Mechanophore Nanoparticles for Deep-Tissue Mechanochemical Dynamic Therapy.

Photodynamic therapy (PDT) and sonodynamic therapy (SDT), using nonionizing light and ultrasound to generate reactive oxygen species, offer promising localized treatments for cancers. However, the effectiveness of PDT is hampered by inadequate tissue penetration, and SDT largely relies on pyrolysis and sonoluminescence, which may cause tissue injury and imprecise targeting. To address these issues, we have proposed a mechanochemical dynamic therapy (MDT) that uses free radicals generated from mechanophore-embedded polymers under mechanical stress to produce reactive oxygen species for cancer treatment. Yet, their application in vivo is constrained by the bulk form of the polymer and the need for high ultrasound intensities for activation. In this study, we developed injectable, nanoscale mechanophore particles with enhanced ultrasound sensitivity by leveraging a core-shell structure comprising silica nanoparticles (NPs) whose interfaces are linked to polymer brushes by an azo mechanophore moiety. Upon focused ultrasound (FUS) treatment, this injectable NP generates reactive oxygen species (ROS), demonstrating promising results in both an in vitro 4T1 cell model and an in vivo mouse model of orthotopic breast cancers. This research offers an alternative therapy technique, integrating force-responsive azo mechanophores and FUS under biocompatible conditions.

Influence of ultrasound-assisted extraction on the pectin extraction yield and structural characteristics: A case study on carrot pomace (Daucus carota)

For the ultrasound-assisted extraction (UAE) of pectin from carrot pomace, the influence of different extraction parameters on the pectin yield and structure was studied. Therefore, an Orthogonal Minimally Aliased Response Surface (OMARS) design and a constant extraction temperature were used. This study showed that pH had the most pronounced effect on the pectin yield and molecular structure, and that a low pH is crucial to liberate pectin from the cell wall matrix. Besides, it was shown that an increased pectin yield can be achieved with an increased ultrasound intensity, provided that the pH is sufficiently low. The optimal UAE conditions resulted in a pectin yield of 57.2 %, while the control extraction, which used the same extraction conditions without the application of ultrasound waves, only resulted in a pectin yield of 37.4 %, showing a significant increase with ultrasonication. Due to the temperature-control during the ultrasonication process, this increased yield can be attributed to the ultrasound waves as such, and not to the accompanying temperature increase. Additionally, shorter extraction times can be used to obtain pectin yields comparable to a conventional acid extraction process. These increased yields and shorter extraction processes show the potential of UAE as an alternative extraction technology.

Removal of Acid Orange 7 dye using Makgeolli lees with ultrasonic assistance

AbstractThis study investigated the efficient removal of Acid Orange 7 (AO7) dye in water by using Makgeolli lees, a popular by‐product obtained during the production of traditional Makgeolli beverages in Korea. By incorporating ultrasound, the effects of contact time, Makgeolli lees dosage, initial AO7 dye concentration, and initial pH of the dye solution were investigated and comprehensively compared with the same experiment using the stirring method. The results consistently showed ultrasound not only enhances the excellent adsorption ability of Makgeolli lees but also accelerates the process compared to the stirring method. The Langmuir isotherm model best described the adsorption process for both methods, suggesting monolayer adsorption on the surface of Makgeolli lees, with maximum capacities of 25.13 mg/g for ultrasound at 40 kHz and 20.41 mg/g for stirring methods. Furthermore, the study showed that optimal dye removal efficiency can be achieved with ultrasound conditions at 28 kHz frequency, 125 W/L power density, and 100% ultrasound intensity. This research promises that the integration of low‐cost biomass coupled with ultrasound could provide a potential solution for dye wastewater treatment.

Bactericidal effect of ultrasound on glutinous rice during soaking and its influence on physicochemical properties of starch and quality characteristics of sweet dumplings

The soaking process of glutinous rice allows the growth and reproduction of microorganisms, which can easily cause food safety problems. In this work, the effects of different ultrasonic powers (150 W, 300 W, 450 W, and 600 W) on the bactericidal effect of glutinous rice, the physicochemical properties of starch and the quality characteristics of sweet dumplings were studied. Compared with soaking for 0 and 2 h, sonication of glutinous rice after soaking for 4 h was more effective at reducing the number of microorganisms in soaked glutinous rice, and the bactericidal effect increased with increasing ultrasound intensity. After 30 min, the total number of bacteria decreased by 2.04 log CFU/g. Moreover, ultrasonic treatment destroys the grain structure of glutinous rice starch, resulting in the formation of dents and cracks on the starch surface, increasing the amylose content, improving its expansion, reducing its short-range order and relative crystallinity, and altering its gelatinization characteristics. In addition, ultrasonic treatment increased the soup transparency of sweet dumplings from 51.8 % to 63.95 %, reducing their hardness, chewiness and adhesiveness. In summary, ultrasonic treatment can not only effectively kill microorganisms in soaked glutinous rice but also improve the quality of glutinous rice dumplings by changing the physicochemical properties of glutinous rice starch. The results of this study provide theoretical support for the application of ultrasonic technology in glutinous rice food production.

Study on ultrasound-enhanced molecular transport in articular cartilage.

Local intra-articular administration with minimal side effects and rapid efficacy is a promising strategy for treating osteoarthritis(OA). Most drugs are rapidly cleared from the joint space by capillaries and lymphatic vessels before free diffusion into cartilage. Ultrasound, as a non-invasive therapy, enhances molecular transport within cartilage through the mechanisms of microbubble cavitation and thermal effects. This study investigated the mass transfer behavior of solute molecules with different molecular weights (479 Da, 40 kDa, 150 kDa) within porcine articular cartilage under low-frequency ultrasound conditions of 40 kHz and ultrasound intensities of 0.189 W/cm2 and 0.359 W/cm2. The results revealed that under the conditions of 0.189 W/cm2 ultrasound intensity, the mass transfer concentration of solute molecules were higher compared to passive diffusion, and with an increase in ultrasound intensity to 0.359 W/cm2, the mass transfer effect within the cartilage was further enhanced. Ultrasound promotes molecular transport in different layers of cartilage. Under static conditions, after 2 h of mass transfer, the concentration of small molecules in the superficial layer is lower than that in the middle layer. After applying ultrasound at 0.189 W/cm2, the molecular concentration in the superficial layer significantly increases. Under conditions of 0.359 W/cm2, after 12 h of mass transfer, the concentration of medium and large molecules in the deep layer region increased by more than two times. In addition, this study conducted an assessment of damage to porcine articular cartilage under ultrasound exposure, revealing the significant potential of low-frequency, low-intensity ultrasound in drug delivery and treatment of OA.

Bioactive, Pro-Apoptotic-Angiotensin Converting Enzyme Inhibitor Effects and Properties of Ultrasound-Treated Traditional Poppy Vinegar Using the Response Surface Methodology Model.

Poppy vinegar with functional properties is a fermented product. This study evaluated traditionally produced poppy vinegar. The study was conducted on poppy vinegar to determine the maximum increase in angiotensin converting enzyme (ACE) inhibitory activity %, total phenolic content (TPC), and radical scavenging activity (DPPH) of the vinegar at different combinations of ultrasound treatment duration (2-14 min) and amplitude (40-100%). The optimal parameters obtained using the response surface methodologies (RSM) were the duration of the ultrasound of 5.5 min and the amplitude of the ultrasound at 57%. When the DPPH values, ACE inhibition %, and TPC and DPPH values obtained with the RSM model were compared with the experimental values, the difference was 9.80, 3.0, and 4.6%, respectively, showing good agreement between actual and predicted values. The higher ultrasound intensities and longer treatment times had a significant effect on antioxidant activity. Poppy vinegar samples significantly induced the apoptosis of lung cancer cells, particularly those stored for 6 and 12 months. The amounts of protocatechuic acid, gallic acid, neohesperidin, hydroxybenzoic acid, resveratrol, rutin, trans-cinnamic acid, quercetin, and flavon in poppy vinegar were determined, which decreased significantly as storage time increased. TPC and TFC were determined to be 90.39 mg of GAE/100 mL and 29.86 mg of TEAC/mL, respectively, and there was no significant change in these bioactive compounds after 6 months of storage. The highest value of ACE inhibitory activity was found at the beginning of the storage period. The present study was the first study to examine the bioactive components, ACE inhibition activity, pro-apoptotic activities, and phenolic composition of traditionally produced ultrasound-treated poppy vinegar during storage. The control of production parameters and the design of ideal poppy vinegar fermentation processes could benefit from this research.

Ultrasound‐Driven Highly Stable Implantable Triboelectric Nanogenerator with Human‐Tissue Acoustic Impedance‐Matched Polyether Ether Ketone

AbstractImplantable electrical neurostimulators offer a promising avenue for treating neurological disorders. However, their dependency on a finite battery life limits their long‐term utility. Emerging transcutaneous ultrasound‐driven triboelectric nanogenerator (TENG) techniques provide solutions for converting external ultrasound waves into internal electricity. This study proposes an implantable ultrasound‐driven TENG (IU‐TENG) using polyether ether ketone (PEEK) for its exceptional stability inside a human body and acoustic impedance compatibility with human tissues. This IU‐TENG remarkably surpasses traditional titanium‐based encapsulation, resulting in a 99.94% efficiency in ultrasound transmission. In addition, PEEK contains numerous electron‐donating functional groups, making it suitable for TENG applications, particularly as a positive triboelectric layer. The device exhibits robust voltage outputs, reaching up to 11.50 and 8.75 V in water and in vivo, respectively, under body‐safe ultrasound intensities. Moreover, its ability to sustain a stable electrical output for over 300 min emphasizes the durability and mechanical resilience of PEEK. In vivo mouse models and ex vivo porcine tissue trials demonstrate the effectiveness of the IU‐TENG in nerve stimulation, showing its potential in medical treatments, enhancing the functionality and longevity of implantable medical devices.

Effects of transcranial magneto-acoustic stimulation on cognitive function and neural signal transmission in the hippocampal CA1 region of mice

As a new means of brain neuroregulation and research, transcranial magneto-acoustic stimulation (TMAS) uses the coupling effect of ultrasound and a static magnetic field to regulate neural activity in the corresponding brain areas. Calcium ions can promote the secretion of neurotransmitters and play a key role in the transmission of neural signals in brain cognition. In this study, to explore the effects of TMAS on cognitive function and neural signaling in the CA1 region of the hippocampus, TMAS was applied to male 2-month-old C57 mice with a magnetic field strength of 0.3 T and ultrasound intensity of 2.6 W/cm2. First, the efficiency of neural signaling in the CA1 region of the mouse hippocampus was detected by fiber photometry. Second, the effects of TMAS on cognitive function in mice were investigated through multiple behavioral experiments, including spatial learning and memory ability, anxiety and desire for novelty. The experimental results showed that TMAS could improve cognitive function in mice, and the efficiency of neural signaling in the CA1 area of the hippocampus was significantly increased during stimulation and maintained for one week after stimulation. In addition, the neural signaling efficiency in the CA1 area of the hippocampus increased in the open field (OF) experiment and recovered after one week, the neural signaling efficiency in the new object exploration (NOE) experiment was significantly enhanced, and the intensity slowed after one week. In conclusion, TMAS enhances cognitive performance and promotes neural signaling in the CA1 region of the mouse hippocampus.

Effect of ultrasound on the activity and structure of polyphenol oxidase purified from mango (Mangifera indica cv. "Xiaotainong")

This study separated, purified, and identified mango polyphenol oxidase (PPO) in Xiaotainong mango, and the inactivation effect and possible inactivation mechanism of mango PPO by ultrasound (US) was investigated. The purified mango PPO, identified as Alfonso mango chloroplast PPO (NCBI number: XP:044462061.1), exhibited specific enzyme activity of 178,192 U/mg, with a yield of 1.2%, which was 9.85-fold higher than crude enzyme solution. Bioinformatics analysis of XP_044462061.1 indicated that the PPO consisted of a stable hydrophilic protein with a molecular weight of 66.77 kDa, an amino acid number of 593, and a structural domain similar to the TYROSINASE_1 (PS00497, tyrosinase) CuA and TYROSINASE_2 (PS00498; tyrosinase) CuB binding regions. The result showed that increased US treatment intensity and time better facilitated PPO inactivation, with a higher US intensity performing better than extended treatment time. US changed and disrupted the secondary and endogenous hydrophobic structures of the mango PPO, altered the exogenous hydrophobicity, and caused PPO molecule aggregation, ultimately leading to PPO inactivation. This article provided a theoretical basis for the application of US technology for processing mango products.

Development of Focused Ultrasound-Assisted Nanoplexes for RNA Delivery.

RNA-based therapeutics, including siRNA, have obtained recognition in recent years due to their potential to treat various chronic and rare diseases. However, there are still limitations to lipid-based drug delivery systems in the clinical use of RNA therapeutics due to the need for optimization in the design and the preparation process. In this study, we propose adaptive focused ultrasound (AFU) as a drug loading technique to protect RNA from degradation by encapsulating small RNA in nanoliposomes, which we term nanoplexes. The AFU method is non-invasive and isothermal, as nanoplexes are produced without direct contact with any external materials while maintaining precise temperature control according to the desired settings. The controllability of sample treatments can be effectively modulated, allowing for a wide range of ultrasound intensities to be applied. Importantly, the absence of co-solvents in the process eliminates the need for additional substances, thereby minimizing the potential for cross-contaminations. Since AFU is a non-invasive method, the entire process can be conducted under sterile conditions. A minimal volume (300 μL) is required for this process, and the treatment is speedy (10 min in this study). Our in vitro experiments with silencer CD44 siRNA, which performs as a model therapeutic drug in different mammalian cell lines, showed encouraging results (knockdown > 80%). To quantify gene silencing efficacy, we employed quantitative polymerase chain reaction (qPCR). Additionally, cryo-electron microscopy (cryo-EM) and atomic force microscopy (AFM) techniques were employed to capture images of nanoplexes. These images revealed the presence of individual nanoparticles measuring approximately 100-200 nm in contrast with the random distribution of clustered complexes observed in ultrasound-untreated samples of liposome nanoparticles and siRNA. AFU holds great potential as a standardized liposome processing and loading method because its process is fast, sterile, and does not require additional solvents.

Controlled ultrasonic interventions through the human skull.

Transcranial focused ultrasound enables precise and non-invasive manipulations of deep brain circuits in humans, promising to provide safe and effective treatments of various neurological and mental health conditions. Ultrasound focused to deep brain targets can be used to modulate neural activity directly or localize the release of psychoactive drugs. However, these applications have been impeded by a key barrier-the human skull, which attenuates ultrasound strongly and unpredictably. To address this issue, we have developed an ultrasound-based approach that directly measures and compensates for the ultrasound attenuation by the skull. No additional skull imaging, simulations, assumptions, or free parameters are necessary; the method measures the attenuation directly by emitting a pulse of ultrasound from an array on one side of the head and measuring with an array on the opposite side. Here, we apply this emerging method to two primary future uses-neuromodulation and local drug release. Specifically, we show that the correction enables effective stimulation of peripheral nerves and effective release of propofol from nanoparticle carriers through an ex vivo human skull. Neither application was effective without the correction. Moreover, the effects show the expected dose-response relationship and targeting specificity. This article highlights the need for precise control of ultrasound intensity within the skull and provides a direct and practical approach for addressing this lingering barrier.

A simulation study of transcranial magnetoacoustic stimulation of the basal ganglia thalamic neural network to improve pathological beta oscillations in Parkinson's disease

BackgroundParkinson's disease (PD) is a common neurodegenerative disease. Transcranial magnetoacoustic stimulation (TMAS) is a new therapy that combines a transcranial focused acoustic pressure field with a magnetic field to excite or inhibit neurons in targeted area, which suppresses the abnormally elevated beta band amplitude in PD states, with high spatial resolution and non-invasively. ObjectiveTo study the effective stimulation parameters of TMAS mononuclear and multinuclear stimulation for the treatment of PD with reduced beta band energy, improved abnormal synchronization, and no thermal damage. MethodsThe TMAS model is constructed based on the volunteer's computed tomography, 128 arrays of phase-controlled transducers, and permanent magnets. A basal ganglia-thalamic (BG-Th) neural network model of the PD state was constructed on the basis of the Izhikevich model and the acoustic model. An ultrasound stimulation neuron model is constructed based on the Hodgkin-Huxley model. Numerical simulations of transcranial focused acoustic pressure field, temperature field and induced electric field at single and dual targets were performed using the locations of STN, GPi, and GPe in the human brain as the main stimulation target areas. And the acoustic and electric parameters at the focus were extracted to stimulate mononuclear and multinuclear in the BG-Th neural network. ResultsWhen the stimulating effect of ultrasound is ignored, TMAS-STN simultaneously inhibits the beta-band amplitude of the GPi nucleus, whereas TMAS-GPi fails to simultaneously have an inhibitory effect on the STN. TMAS-STN&GPi can reduce the beta band amplitude. TMAS-STN&GPi&GPe suppressed the PD pathologic beta band amplitude of each nucleus to a greater extent. When considering the stimulatory effect of ultrasound, lower sound pressures of ultrasound do not affect the neuronal firing state, but higher sound pressures may promote or inhibit the stimulatory effect of induced currents. ConclusionsAt 9 T static magnetic field, 0.5–1.5 MPa and 1.5–2.0 MPa ultrasound had synergistic effects on individual STN and GPi neurons. TMAS multinuclear stimulation with appropriate ultrasound intensity was the most effective in suppressing the amplitude of pathological beta oscillations in PD and may be clinically useful.

Effect of Some Technological Factors of Extraction on Total Lentinan Content in Sapa Shiitake Mushroom Extract

In Vietnam, the source of shiitake mushroom is very abundant and grown in many provinces/cities, the total yield reaches hundreds of thousands of tons/per year, mainly serving the demand of domestic food processing, and did not develop into medicinal mushrooms yet. Among them, Sapa shiitake mushroom is being widely cultivated in Sapa-Vietnam, and is a raw material with great potential for lentinan exploitation. Until now, researches on extracting and obtaining lentinan in Vietnam are still limited. Those are reasons to carry out this research. The effects of solvent in combination with assistance of ultrasound wave in lentinan extracting capability in Sapa shiitake mushroom were studied. Before carrying out extracting of lentinan; The dried fruit bodies of Sapa shiitake mushroom was crushed in to 1mm. Five extracting parameters include concentration of Na0H solvent (%), proportion of Na0H solvent and raw material (v:w), extracting temperature, ultrasound time and ultrasound intensity were carried out. An extraction without the use of ultrasound for 180 min was control sample. Total lentinan content was obtained during the extracting process. The results indicated that shiitake mushrooms were extracted by using 0.35% Na0H solvent, the rate of Na0H solvent and mushrooms was 12:1 (v:w), intensity of ultrasound was 58 W/cm2, frequency was 20 kHz, extracting temperature was 65 &amp;deg;C, time of extraction was 6 min gave total lentinan content 1.63 times higher than the control.