Lentil protein isolate (Lens culinaris) subjected to ultrasound treatment combined or not with heat-treatment: structural characterization and ability to stabilize high internal phase emulsions.

This study evaluated the effect of ultrasound treatment combined or not with heat treatment applied to lentil protein isolate (LPI) aiming to enhance its ability to stabilize high internal phase emulsions (HIPE). LPI dispersion (2%, w/w) was ultrasound-treated at 60% (UA) and 70% (UB) amplitude for 7 min; these samples were subjected to and then heat treatments at 70 °C (UAT70 and UBT70, respectively) or 80 °C (UAT80 and UBT80, respectively) for 20 min. HIPEs were produced with 25% untreated and treated LPI dispersions and 75% soybean oil using a rotor-stator (15,500 rpm/1 min). The LPI dispersions were evaluated for particle size, solubility, differential scanning calorimetry, electrophoresis, secondary structure estimation (circular dichroism and FT-IR), intrinsic fluorescence, surface hydrophobicity, and free sulfhydryl groups content. The HIPEs were evaluated for droplet size, morphology, rheology, centrifugal stability, and the Turbiscan test. Ultrasound treatment decreased LPI dispersions' particle size (∼80%) and increased solubility (∼90%). Intrinsic fluorescence and surface hydrophobicity confirmed LPI modification due to the exposure to hydrophobic patches. The combination of ultrasound and heat treatments resulted in a reduction in the free sulfhydryl group content of LPI. HIPEs produced with ultrasound-heat-treated LPI had a lower droplet size distribution mode, greater oil retention values in the HIPE structure (> 98%), lower Turbiscan stability index (< 2), and a firmer and more homogeneous appearance compared to HIPE produced with untreated LPI, indicating higher stability for the HIPEs stabilized by treated LPI. Therefore, combining ultrasound and heat treatments could be an effective method for the functional modification of lentil proteins, allowing their application as HIPE emulsifiers.

Tetracycline degradation by dual-frequency ultrasound combined with peroxymonosulfate.

Tetracycline has received a great deal of interest for the harmful effects of substance abuse on ecosystems and humanity. The effects of different processes on the degradation of tetracycline were compared, with dual-frequency ultrasound (DFUS) in combination with peroxymonosulfate (PMS) being the most effective for the tetracycline degradation. Free radical scavenging experiments showed that O2∙-,SO4∙- and •OH were the main reactive radicals in the degradation of tetracycline. According to the major intermediates of tetracycline degradation identified, three possible degradation pathways were proposed, which are of significance for translational studies of tetracycline degradation. Notably, these intermediates were found to be significantly less toxicity. The number of active bubbles in the degradation vessel was calculated using a semi-empirical formula, and a higher value of 1.44 × 108 L-1s-1 of bubbles was obtained when using dual-frequency ultrasound at 20 kHz (210 W/L) and 80 kHz (85.4 W/L). Therefore, compared to 20 kHz, although the yield of strong oxidizing substances from individual active bubbles decreased slightly, a significant increment of the number of active bubbles still resulted in a higher synergistic effect, and the combination of DFUS and PMS should be effective in promoting the generation of reactive free radicals and mass transfer processes within the degradation vessel, which provides a method for efficient removal of tetracycline from wastewater.

Ultrasonication followed by aqueous two-phase system for extraction, on-site modification and isolation of microalgal starch with reduced digestibility.

Microalgae are new and sustainable sources of starch with higher productivity and flexible production modes than conventional terrestrial crops, but the downstream processes need further development. Here, ultrasonication (with power of 200 W or 300 W and duration of 10, 15, 20, or 25 min) was applied to simultaneously extract and modify starch from a marine microalga Tetraselmis subcordiformis for reducing the digestibility, and an aqueous two-phase system (ATPS) of ethanol/NaH2PO4 was then used to isolate the starches with varied properties. Increasing ultrasonic duration facilitated the partition of starch into the bottom pellet, while enhancing the ultrasonic power was conducive to the allocation in the interphase of the ATPS. The overall starch recovery yield reached 73 âˆ¼ 87 % and showed no significant difference among the ultrasonic conditions tested. The sequential ultrasonication-ATPS process successfully enriched the starch with purities up to 65 % âˆ¼ 88 %, which was among the top levels reported in microalgal starch isolated. Ultrasonication produced more amylose which was mainly fractionated into the interface of the ATPS. The digestibility of the starch was altered under different ultrasonic conditions and varied from different ATPS phases as well, with the one under the ultrasonic power of 200 W for 15 min at the bottom pellet having the highest resistant starch content (RS, 39.7 %). The structural and compositional analysis evidenced that the ultrasonication-ATPS process could exert impacts on the digestibility through altering the surface roughness and fissures of the starch granules, modulating the impurity compositions (protein and lipid) that could interact with starch, and modifying the long- and short-range ordered structures. The developed ultrasonication-ATPS process provided novel insights into the mechanism and strategy for efficient production of functional starch from microalgae with a potential in industrial application.

Ultrasound assisted extraction of phytochemicals from Piper betel L.

Piper betel contains phytochemicals with diverse pharmacological effects. The objective of this study was to enhance the extraction efficiency of phytochemicals and the chlorophyll content using ultrasonication. The Box-Behnken design was employed to optimize the time (10, 20, 30 min), temperature (20, 30, and 40 °C), and solid-solvent ratio (1:10, 1:20, 1:30) by utilizing response surface methods with three independent variables. Multiple parameters, including extract yield, total phenol, total flavonoid, antioxidant activity, and chlorophyll content were used to optimize the conditions. The linear relationship between power intensity and responses was determined to be statistically significant, with a p-value less than 0.01. The interaction effect of temperature, time, and ratio of solid solvent was shown to be statistically significant (p < 0.05) for all the obtained results. The optimal parameters for achieving the highest extract yield were as follows: a temperature of 40 °C, a sonication time of 30 min, and a solid solvent ratio of 1:10. These conditions result in an extract yield of 21.99 %, a total flavonoid content of 44.97 mg/GAE, a total phenolic content of 185.05 mg/GAE, a DPPH scavenging activity of 99.1 %, and a chlorophyll content of 49.95 mg/ml. This study highlights the significance of customized extraction methodologies for optimizing the bioactive capacity of phytochemicals derived from betel leaves. The elucidation of extraction parameters and the resultant phytochemical profiles serves as a fundamental framework for the advancement of innovative pharmaceuticals and nutraceuticals, capitalizing on the therapeutic attributes of this traditional medicinal botanical.

DES-ultrasonication treatment of cellulose nanocrystals and the reinforcement in carrageenan biocomposite.

CNCs are intensively studied to reinforce biocomposites. However, it remains a challenge to homogeneously disperse the CNC in biocomposites for a smooth film surface. Mechanochemical treatment via ultrasonication in deep eutectic solvent (DES) generated a stable dispersion of CNC before incorporation into carrageenan biocomposite. Shifted peaks of choline chloride (ChCl) methylene groups to 3.95-3.98 ppm in 1H NMR indicated a formation of eutectic mixture between the hydrogen bond acceptor (HBA) and hydrogen bond donor (HBD) at the functional group of CH3···OH. The swelling of CNC in the DES was proven by the formation of intermolecular H-bond at a length of 2.46 Å. The use of DES contributed to a good dispersion of CNC in the solution which increased zeta potential by 43.2 % compared to CNC in deionized water. The ultrasonication amplitude and feed concentration were varied for the best parameters of a stable dispersion of CNC. The crystallinity of 1 wt% of CNC at 20 % sonication amplitude improved from 76 to 81 %. The high crystallinity of CNCDES resulted in an increase in film tensile and capsule loop strength of Carra-CNCDES by 20.7 and 19.4 %, respectively. Improved dispersion of CNCDES reduced the surface roughness of the biocomposite by 21.8 %. H-bond network in CNCDES improved the biocomposite properties for an ingenious reinforcement material.

Development and application of a protocol for extractables profiling from central venous catheters in neonates.

Peripherally inserted central catheters (PICC-lines) used in neonatology are made of thermoplastic polyurethane (TPU) or silicone. These materials usually contain substances that may leach into drug vehicles or blood. In this extractables study, we determined the optimal extraction conditions using TPU films containing defined amounts of butylhydroxytoluene (BHT) and then applied them on unused and explanted PICC-lines. Maceration and sonication tests were carried out with hexane, acetone and water as the extraction solvents. The analyses were performed using gas and liquid chromatography coupled with mass spectrometry detectors, as well as inductive coupled plasma optical emission spectroscopy to detect a wide range of extractables. We selected a limited list of substances to be sought from the usual adjuvants and monomers, related to their carcinogenic, mutagenic or reprotoxic properties and/or existence in endocrine disruptors lists. The TPU-film experiments showed that acetone was slightly better than hexane, and maceration better than sonication. When applied to PICC-lines, the extraction methods were almost similar but acetone was clearly better than hexane for TPU. From the 48 peaks initially observed in GC-MS, we ended up with 37 peaks to follow in TPU PICC-lines, among which were those of BHT and 4,4'-Methylenebis(cyclohexyl isocyanate) isomers. For silicone PICC-lines, out of 41 peaks initially observed in GC-MS, we followed 20 peaks, most of them being identified as cyclosiloxanes. Barium was the main inorganic element extracted for both PICC-lines. For TPU PICC-lines, the inter-batch variability was higher than for intra-batch, but in silicone devices both were similar. When compared to new PICC-lines, explanted TPU PICC-lines extracted peaks had a lower area under the curve (AUC), while the AUCs of the peaks were higher for the majority of silicone PICC-lines extract compounds. No identified substances were detected above their toxicological threshold, but isocyanates and cyclosiloxanes toxicity was mostly studied for other exposition routes than intravenous. The methods defined in this study were efficient in producing extractable profiles from both PICC-lines.

Thermosonication as an effective substitution for fusion in Brazilian cheese spread (Requeijão Cremoso) manufacturing: The effect of ultrasonic power on technological properties.

In this initial study, the impact of thermosonication as an alternative to the traditional fusion in Brazilian cheese spread (Requeijão Cremoso) manufacture was investigated. The effect of ultrasound (US) power was evaluated considering various aspects such as gross composition, microstructure, texture, rheology, color, fatty acid composition, and volatile compounds. A 13 mm US probe operating at 20 kHz was used. The experiment involved different US power levels (200, 400, and 600 W) at 85 °C for 1 min, and results were compared to the conventional process in the same conditions (85 °C for 1 min, control treatment). The texture became softer as ultrasound power increased from 200 to 600 W, which was attributed to structural changes within the protein and lipid matrix. The color of the cheese spread also underwent noticeable changes for all US treatments, and treatment at 600 W resulted in increased lightness but reduced color intensity. Moreover, the fatty acid composition of the cheese spread showed variations with different US power, with samples treated at 600 W showing lower concentrations of saturated and unsaturated fatty acids, as well as lower atherogenicity and thrombogenicity indexes, indicating a potentially healthier product. Volatile compounds were also influenced by US, with less compounds being identified at higher powers, especially at 600 W. This could indicate possible degradation, which should be evaluated in further studies regarding US treatment effects on consumer perception. Hence, this initial work demonstrated that thermosonication might be interesting in the manufacture of Brazilian cheese spread, since it can be used to manipulate the texture, color and aroma of the product in order to improve its quality parameters.

Coffee brewing sonoreactor for reducing the time of cold brew from several hours to minutes while maintaining sensory attributes.

This research designed and developed an ultrasonic reactor for a fast and on demand production of cold brew coffee, remarkably reducing the brewing time from 24 h to less than 3 min. The technology was engineered by utilizing resonance to induce ultrasonic waves around the walls of the brewing basket of an espresso machine. The sound transmission system comprised a transducer, a horn and a brewing basket. This arrangement transformed the coffee basket into an effective sonoreactor that injected sound waves at multiple points through its walls, thereby generating multiple regions for acoustic cavitation within the reactor. Furthermore, acoustic streaming induced greater mixing and enhanced mass transfer during brewing. The design was accomplished by modeling the transmission of sound, and acoustic cavitation. Brew characterization and chemical composition analysis was performed, considering factors such as pH, acidity, color, and the composition of caffeine, fatty acids, and volatiles. The efficiency of the extraction increased by decreasing the basket loading percentage (BLP). For instance, sonicating at 100 W doubled the extraction yield and caffeine concentration, from 15.05 % to 33.44 % at BLP = 33 %, and from 0.91 mg/mL to 1.84 mg/mL at BLP = 67 %, respectively. The total fatty acids increased from 1.16 mg/mL to 9.20 mg/mL, representing an eightfold increase, at BLP = 33 %. Finally, a sensory analysis was conducted to evaluate appearance, aroma, texture, flavor, and aftertaste, which demonstrated that coffee brewed for 1 and 3 min in the sonoreactor exhibited almost undistinguishable properties compared to a standard 24 h brewing without ultrasound.

Effects of different sonication parameters of theta burst transcranial ultrasound stimulation on human motor cortex.

BACKGROUND: Theta burst TUS (tbTUS) can induce increased cortical excitability in human, but how different sonication parameters influence the effects are still unknown. OBJECTIVE: To examine how a range of sonication parameters, including acoustic intensity, pulse repetition frequency, duty cycle and sonication duration, influence the effects of tbTUS on human motor cortical excitability. METHODS: 14 right-handed healthy subjects underwent 8 sessions with different tbTUS parameters in a randomized, cross-over design on separate days. The original tbTUS protocol was studied in one session and one parameter was changed in each of the seven sessions. To examine changes in cortical excitability induced by tbTUS, we measured the motor-evoked potential (MEP) amplitude, resting motor threshold, short-interval intracortical inhibition and intracortical facilitation, as well as short-interval intracortical facilitation before and up to 90 min after tbTUS. RESULTS: All conditions increased MEP amplitudes except the condition with low acoustic intensity of 10 W/cm2. Pulse repetition frequency of 5 Hz produced higher MEP amplitudes compared to pulse repetition frequencies of 2 and 10 Hz. In addition, higher duty cycles (5%, 10%, and 15%) and longer sonication durations (40, 80, and 120 s) were associated with longer duration of increased MEP amplitudes. Resting motor threshold remained stable in all conditions. For paired-pulse TMS measures, tbTUS reduced short-interval intracortical inhibition and enhanced short-interval intracortical facilitation, but had no effect on intracortical facilitation. CONCLUSIONS: Ultrasound bursts repeated at theta (∼5 Hz) frequency is optimal to produce increased cortical excitability with the range of 2-10 Hz. Furthermore, there was a dose-response effect regarding duty cycle and sonication duration in tbTUS for plasticity induction. The aftereffects of tbTUS were associated with a shift of the inhibition/excitation balance toward less inhibition and more excitation in the motor cortex. These findings can be used to determine the optimal tbTUS parameters in neuroscience research and treatment of neurological and psychiatric disorders.

Nanobubble-mediated cancer cell sonoporation using low-frequency ultrasound.

Ultrasound insonation of microbubbles can form transient pores in cell membranes that enable the delivery of non-permeable extracellular molecules to the cells. Reducing the size of microbubble contrast agents to the nanometer range could facilitate cancer sonoporation. This size reduction can enhance the extravasation of nanobubbles into tumors after an intravenous injection, thus providing a noninvasive sonoporation platform. However, drug delivery efficacy depends on the oscillations of the bubbles, the ultrasound parameters and the size of the target compared to the membrane pores. The formation of large pores is advantageous for the delivery of large molecules, however the small size of the nanobubbles limit the bioeffects when operating near the nanobubble resonance frequency at the MHz range. Here, we show that by coupling nanobubbles with 250 kHz low frequency ultrasound, high amplitude oscillations can be achieved, which facilitate low energy sonoporation of cancer cells. This is beneficial both for increasing the uptake of a specific molecule and to improve large molecule delivery. The method was optimized for the delivery of four fluorescent molecules ranging in size from 1.2 to 70 kDa to breast cancer cells, while comparing the results to targeted microbubbles. Depending on the fluorescent molecule size, the optimal ultrasound peak negative pressure was found to range between 300 and 500 kPa. Increasing the pressure to 800 kPa reduced the fraction of fluorescent cells for all molecules sizes. The optimal uptake for the smaller molecule size of 4 kDa resulted in a fraction of 19.9 ± 1.8% of fluorescent cells, whereas delivery of 20 kDa and 70 kDa molecules yielded 14 ± 0.8% and 4.1 ± 1.1%, respectively. These values were similar to targeted microbubble-mediated sonoporation, suggesting that nanobubbles can serve as noninvasive sonoporation agents with a similar potency, and at a reduced bubble size. The nanobubbles effectively reduced cell viability and may thus potentially reduce the tumor burden, which is crucial for the success of cancer treatment. This method provides a non-invasive and low-energy tumor sonoporation theranostic platform, which can be combined with other therapies to maximize the therapeutic benefits of cancer treatment or be harnessed in gene therapy applications.

Small volume blood-brain barrier opening in macaques with a 1 MHz ultrasound phased array.

The use of focused ultrasound to open the blood-brain barrier (BBB) has the potential to deliver drugs to specific regions of the brain. The size of the BBB opening and ability to localize the opening determines the spatial extent and is a limiting factor in many applications of BBB opening where targeting a small brain region is desired. Here we evaluate the performance of a system designed for small opening volumes and highlight the unique challenges associated with pushing the spatial precision of this technique. To achieve small volume openings in cortical regions of the macaque brain, we tested a custom 1 MHz array transducer integrated into a magnetic resonance image-guided focused ultrasound system. Using real-time cavitation monitoring, we demonstrated twelve instances of single sonication, small volume BBB opening with average volumes of 59 ± 37 mm3 and 184 ± 2 mm3 in cortical and subcortical targets, respectively. We found high correlation between subject-specific acoustic simulations and observed openings when incorporating grey matter segmentation (R2 = 0.8577), and the threshold for BBB opening based on simulations was 0.53 MPa. Analysis of MRI-based safety assessment and cavitation signals indicate a safe pressure range for 1 MHz BBB opening and suggest that our system can be used to deliver drugs and gene therapy to small brain regions.

Influence of focused ultrasound on locoregional drug delivery to the brain: Potential implications for brain tumor therapy.

INTRODUCTION: Efficient delivery of therapeutics across the blood-brain barrier (BBB) for the treatment of central nervous system (CNS) tumors is a major challenge to the development of safe and efficacious therapies. Locoregional drug delivery platforms offer an improved therapeutic index by achieving high drug concentrations in the target tissue with negligible systemic exposure. Intrathecal (intraventricular) [IT] and convection-enhanced delivery [CED] are two clinically relevant methods being employed for various CNS malignancies. Both of these standalone platforms suffer from passive post-administration distribution forces, sometimes limiting the desired distribution for tumor therapy. Focused ultrasound and microbubble-mediated blood-brain barrier opening (FUS-BBBO) is a recent modality used for enhanced drug delivery. It is postulated that coupling of FUS with these alternative delivery routes may provide benefits. Multimodality FUS may provide the desired ability to increase the depth of parenchymal delivery following IT administration and provide a means for contour directionality with CED. Further, the transient enhanced permeability achieved with FUS-BBBO is well established, but drug residence and transit times, important to clinical dose scheduling, have not yet been defined. The present investigation comprises two discrete studies: 1. Conduct a comprehensive quantitative evaluation to elucidate the effect of FUS-BBBO as it relates to varying routes of administration (IT and IV) in its capacity to facilitate drug penetration within the striatal-thalamic region. 2. Investigate the impact of combining FUS-BBBO with CED on drug distribution, with a specific focus on the temporal dynamics of drug retention within the target region. METHODS: Firstly, we quantitatively assessed how FUS-BBBO coupled with IT and IV altered fluorescent dye (Dextran 2000 kDa and 70 kDa) distribution and concentration in a predetermined striatal-thalamic region in naïve mice. Secondly, we analyzed the pharmacokinetic effects of using FUS mediated BBB disruption coupled with CED by measuring the volume of distribution and time-dependent concentration of the dye. RESULTS: Our results indicate that IV administration coupled with FUS-BBBO successfully enhances delivery of dye into the pre-defined sonication targets. Conversely, measurable dye in the sonication target was consistently less after IT administration. FUS enhances the distribution volume of dye after CED. Furthermore, a shorter time of residence was observed when CED was coupled with FUS-BBBO application when compared to CED alone. CONCLUSION: 1. Based on our findings, IV delivery coupled with FUS-BBBO is a more efficient means for delivery to deep targets (i.e. striatal-thalamic region) within a predefined spatial conformation compared to IT administration. 2. FUS-BBBO increases the volume of distribution (Vd) of dye after CED administration, but results in a shorter time of residence. Whether this finding is reproducible with other classes of agents (e.g., cytotoxic agents, antibodies, viral particles, cellular therapies) needs to be studied.

Complex Dependence of Escherichia coli-based Cell-Free Expression on Sonication Energy During Lysis.

Cell lysis─by sonication or bead beating, for example─is a key step in preparing extracts for cell-free expression systems. To create high protein-production capacity extracts, standard practice is to lyse cells sufficiently to thoroughly disrupt the membrane and thus extract expression machinery but without degrading that machinery. Here, we investigate the impact of different sonication energy inputs on the protein-production capacity of Escherichia coli extracts. While the existence of operator-specific optimal sonication energy inputs is widely known, our findings show that the sonication energy input that yields maximal protein output from a given expression template may depend on plasmid concentration, transcriptional and translational features (e.g., promoter), and other expression vector components (e.g., origin of replication). These results indicate that sonication protocols cannot be standardized to a single optimum, suggest strategies for improving protein yields, and more broadly highlight the need for better metrics and protocols for characterizing cell extracts.

Transcranial focused ultrasound stimulation of cortical and thalamic somatosensory areas in human.

The effects of transcranial focused ultrasound (FUS) stimulation of the primary somatosensory cortex and its thalamic projection (i.e., ventral posterolateral nucleus) on the generation of electroencephalographic (EEG) responses were evaluated in healthy human volunteers. Stimulation of the unilateral somatosensory circuits corresponding to the non-dominant hand generated EEG evoked potentials across all participants; however, not all perceived stimulation-mediated tactile sensations of the hand. These FUS-evoked EEG potentials (FEP) were observed from both brain hemispheres and shared similarities with somatosensory evoked potentials (SSEP) from median nerve stimulation. Use of a 0.5 ms pulse duration (PD) sonication given at 70% duty cycle, compared to the use of 1 and 2 ms PD, elicited more distinctive FEP peak features from the hemisphere ipsilateral to sonication. Although several participants reported hearing tones associated with FUS stimulation, the observed FEP were not likely to be confounded by the auditory sensation based on a separate measurement of auditory evoked potentials (AEP) to tonal stimulation (mimicking the same repetition frequency as the FUS stimulation). Off-line changes in resting-state functional connectivity (FC) associated with thalamic stimulation revealed that the FUS stimulation enhanced connectivity in a network of sensorimotor and sensory integration areas, which lasted for at least more than an hour. Clinical neurological evaluations, EEG, and neuroanatomical MRI did not reveal any adverse or unintended effects of sonication, attesting its safety. These results suggest that FUS stimulation may induce long-term neuroplasticity in humans, indicating its neurotherapeutic potential for various neurological and neuropsychiatric conditions.

Global sonication of the human intracranial space via a jumbo planar transducer.

Contrary to conditioning a Focused Ultrasound (FUS) beam to sonicate a localized region of the human brain, the goal of this investigation was to explore the prospect of distributing homogeneous ultrasound energy over the entire brain space with a large cranium-wide ultrasound beam. Recent ultrasound preclincal studies utilizing large or whole brain stimulation regions create a demand for expanding the treatment envelope of transcranial pulsed-low intensity ultrasound towards Global Brain Sonication (GBS) for potential human investigation. Here, we conduct ultrasound field characterizations when transmitting pulsed ultrasound through human skull specimens using a 1-3 piezocomposite planar transducer operating at 464 kHz with an active single-element surface of 30 × 30 cm. Through computational simulation and hydrophone scanning methodology, ultrasound wave behavior and dose homogeneity in the brain space were evaluated under various trajectories of sonication using the planar transducer. Clinically relevant pulse parameters used for transcranial therapeutic ultrasound applications were used in the experiments. Simulations and empirical testing revealed that dose homogeneity and acoustic intensity over the brain space are influenced by sonication trajectory, skull lens effects, and acoustic wave reflections. The transducer can emit a spatial peak pulse average intensity of 4.03 W/cm2 (0.24 MPa) measured in the free-field at 464 kHz with electrical power of 1 kW. The simulation showed that approximately 99 % of the cranial volume was exposed with <30 % of the maximum external acoustic intensity being transmitted into the skull. The transmission loss across all sonication trajectories is similar to previously reported FUS studies. A marker for GBS dose homogeneity is introduced to score the ultrasound pressure field uniformity in the intracranial space. Results of this study identify the initial challenges of exposing the entire human brain space with ultrasound using a large cranium-wide sonication beam intended for global brain therapeutic modulation.

Newly Designed 3D-Printed Sonication Test Cell Optimized for In Vitro Sonication Experiments.

OBJECTIVE: Precise control over the ultrasound field parameters experienced by biological samples during sonication experiments in vitro may be quite challenging. The main goal of this work was to outline an approach to construction of sonication test cells that would minimize the interaction between the test cells and ultrasound. METHODS: Optimal dimensions of the test cell were determined through measurements conducted in a water sonication tank using 3D-printed test objects. The offset of local acoustic intensity variability inside the sonication test cell was set to value of ±50% of the reference value (i.e., local acoustic intensity measured at last axial maximum in the free-field condition). The cytotoxicity of several materials used for 3D printing was determined using the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay. RESULTS: The sonication test cells were 3D printed from polylactic acid material, which was not toxic to the cells. Silicone membrane HT-6240, which was used to construct the bottom of the test cell, was found to reduce ultrasound energy minimally. Final ultrasound profiles inside the sonication test cells indicated the desired variability of local acoustic intensity. The cell viability in our sonication test cell was comparable to that of commercial culture plates with bottoms constructed with silicone membrane. CONCLUSION: An approach to construction of sonication test cells minimizing the interaction of the test cell and ultrasound has been outlined.

Sustainable emerging high-intensity sonication processing to enhance the protein bioactivity and bioavailability: An updated review.

High-intensity ultrasound (HIU) is considered one of the promising non-chemical eco-friendly techniques used in food processing. Recently (HIU) is known to enhance food quality, extraction of bioactive compounds and formulation of emulsions. Various foods are treated with ultrasound, including fats, bioactive compounds, and proteins. Regarding proteins, HIU induces acoustic cavitation and bubble formation, causing the unfolding and exposure of hydrophobic regions, resulting in functional, bioactive, and structural enhancement. This review briefly portrays the impact of HIU on the bioavailability and bioactive properties of proteins; the effect of HIU on protein allergenicity and anti-nutritional factors has also been discussed. HIU can enhance bioavailability and bioactive attributes in plants and animal-based proteins, such as antioxidant activity, antimicrobial activity, and peptide release. Moreover, numerous studies revealed that HIU treatment could enhance functional properties, increase the release of short-chain peptides, and decrease allergenicity. HIU could replace the chemical and heat treatments used to enhance protein bioactivity and digestibility; however, its applications are still on research and small scale, and its usage in industries is yet to be implemented.

Sonication as a tool for disrupting biofilms and recovering microorganisms in bladder catheters.

INTRODUCTION: Urinary catheter-related infection is commonly associated with bacterial biofilm. The impact of anaerobes is unknown, but their detection in the biofilm on this device has not been previously reported. This study aimed to evaluate the capability to recovery strict, facultative, and aerobic microorganisms in patients using bladder catheters from ICUs using conventional culture, sonication, urinary analysis, and mass spectrometry. METHODS: Parallel, sonicated bladder catheters from 29 critically ill patients were compared with their routine urine culture. Identification was performed using matrix-assisted laser desorption/ionization with time-of-flight mass spectrometry. RESULTS: The positivity rate in urine (n = 2, 3.4%) was lower than that in sonicated catheters (n = 7, 13.8%). CONCLUSION: Bladder catheter sonication showed more positive culture results than urine samples for anaerobic and aerobic microorganisms. The role of anaerobes in urinary tract infection and catheter biofilm is discussed.

Fluid flow influences ultrasound-assisted endothelial membrane permeabilization and calcium flux.

The local fluid dynamics experienced by circulating microbubbles vary across different anatomical sites, which can influence ultrasound-mediated therapeutic delivery efficacy. This study aimed to elucidate the effect of fluid flow rate in combination with repeated short-pulse ultrasound on microbubble-mediated endothelial cell permeabilization. Here, a seeded monolayer of human umbilical (HUVEC) or brain endothelial cells (HBEC-5i) was co-perfused with a solution of microbubbles and propidium iodide (PI) at either a flow rate of 5 or 30 ml/min. Using an acoustically coupled inverted microscope, cells were exposed to 1 MHz ultrasound with 20-cycle bursts, 1 ms PRI, and 2 s duration at a peak negative pressure of 305 kPa to assess the role of flow rate on ultrasound-stimulated endothelial cell permeability, as well as Ca2+ modulation. In addition, the effect of inter-pulse delays (∆t = 1s) on the resulting endothelial permeability was investigated. Our results demonstrate that under an identical acoustic stimulus, fast-flowing microbubbles resulted in a statistically significant increase in cell membrane permeability, at least by 2.3-fold, for both endothelial cells. Likewise, there was a substantial difference in intracellular Ca2+ levels between the two examined flow rates. In addition, multiple short pulses rather than a single pulse ultrasound, with an equal number of bursts, significantly elevated endothelial cell permeabilization, at least by 1.4-fold, in response to ultrasound-stimulated microbubbles. This study provides insights into the design of optimal, application-dependent pulsing schemes to improve the effectiveness of ultrasound-mediated local therapeutic delivery.

Diagnostic accuracy of sonication fluid cultures from prosthetic components in periprosthetic joint infection: an updated diagnostic meta-analysis.

BACKGROUND: Periprosthetic joint infection (PJI) is the most serious complication following total joint arthroplasty (TJA) and has a significant impact on patients and the national healthcare system. To date, the diagnosis of PJI is still confronted with dilemmas. The present study investigated the validity of sonication fluid culture (SFC) for removing implants in the diagnosis of PJI after joint replacement. METHODS: From database establishment to December 2020, relevant literature was retrieved from the PubMed, Web of Science, Embase and Cochrane Library databases. Two reviewers independently performed quality assessment and data extraction to calculate the pooled sensitivity, specificity, positive likelihood ratio (PLR), negative likelihood ratio (NLR), area under the curve (AUC) and diagnostic odds ratio (DOR) to evaluate the diagnostic value of overall SFC for PJI. RESULTS: A total of 38 eligible studies including 6302 patients were selected in this study. The pooled sensitivity, specificity, PLR, NLR, and DOR of SFC for PJI diagnosis were 0.77 (95% confidence interval [CI], 0.76-0.79), 0.96 (95% CI, 0.95-0.96), 18.68 (95% CI, 11.92-29.28), 0.24 (95% CI, 0.21-0.29), and 85.65 (95% CI, 56.46-129.94), respectively, while the AUC was 0.92. CONCLUSION: This meta-analysis showed that SFC was of great value in PJI diagnosis, and the evidence of SFC on PJI was more favorable but not yet strong. Therefore, improvement of the diagnostic accuracy of SFC is still necessary, and the diagnosis of PJI continues to warrant a multiplex approach before and during a revision procedure.