Non-cell-autonomous disruption of nuclear architecture as a potential cause of COVID-19-induced anosmia.

SARS-CoV-2 infects less than 1% of cells in the human body, yet it can cause severe damage in a variety of organs. Thus, deciphering the non-cell-autonomous effects of SARS-CoV-2 infection is imperative for understanding the cellular and molecular disruption it elicits. Neurological and cognitive defects are among the least understood symptoms of COVID-19 patients, with olfactory dysfunction being their most common sensory deficit. Here, we show that both in humans and hamsters, SARS-CoV-2 infection causes widespread downregulation of olfactory receptors (ORs) and of their signaling components. This non-cell-autonomous effect is preceded by a dramatic reorganization of the neuronal nuclear architecture, which results in dissipation of genomic compartments harboring OR genes. Our data provide a potential mechanism by which SARS-CoV-2 infection alters the cellular morphology and the transcriptome of cells it cannot infect, offering insight to its systemic effects in olfaction and beyond.

Exploring the Potential of Ultrasound Therapy to Reduce Skin Scars: An In Vitro Study Using a Multi-Well Device Based on Printable Piezoelectric Transducers.

Excessive skin scarring affects over 100 million patients worldwide, with effects ranging from cosmetic to systemic problems, and an effective treatment is yet to be found. Ultrasound-based therapies have been used to treat a variety of skin disorders, but the exact mechanisms behind the observed effects are still unclear. The aim of this work was to demonstrate the potential of ultrasound for the treatment of abnormal scarring by developing a multi-well device based on printable piezoelectric material (PiezoPaint™). First, compatibility with cell cultures was evaluated using measurements of heat shock response and cell viability. Second, the multi-well device was used to treat human fibroblasts with ultrasound and quantify their proliferation, focal adhesions, and extracellular matrix (ECM) production. Ultrasound caused a significant reduction in fibroblast growth and ECM deposition without changes in cell viability or adhesion. The data suggest that these effects were mediated by nonthermal mechanisms. Interestingly, the overall results suggest that ultrasound treatment would a be beneficial therapy for scar reduction. In addition, it is expected that this device will be a useful tool for mapping the effects of ultrasound treatment on cultured cells.

Lead-Free HIFU Transducers.

High-intensity focused ultrasound transducers operating at 4 MHz based on lead-free piezoceramics from the sodium bismuth titanate (NBT) family are described. First, the piezoelectric material (Pz12X) is evaluated from the standpoint of transducer design and its important characteristics, including temperature dependance of several parameters such as dielectric and mechanical coefficients. Then, the performance of six transducers of the same design is evaluated in terms of electro-acoustic efficiency and its dependency on the operating acoustic power level up to 30 W. Overall, the initial electro-acoustic efficiency of three independent transducers is approximately 50% at low acoustic power levels and slightly drops down to 42% as the input electric power reaches 10 W. This process is stable and fully reversible. Moreover, the stability of electro-acoustic efficiency over extended power burst cycling is studied using another two transducers up to 95 × 103 power bursts of 250-ms duration and acoustic power of 10 W. This protocol is beyond the typical clinical use of similar devices in practice. No significant changes in electro-acoustic performance are noted. Additionally, the input electric power and the output acoustic power, together with the temperature of the piezoelectric component, are evaluated simultaneously over the period of one power burst. It is found that the maximum operating temperature over a high-input electric power burst of 600 J is below 60°C, which defines the operational limit for such devices, as the de-poling temperature of the lead-free material is around 85°C. It is found that the lead-free material from the NBT family is also a promising alternative to lead-based PZT-type materials in high-power therapeutic ultrasound.

The Combination of Low-Frequency Ultrasound and Antibiotics Improves the Killing of In Vitro Staphylococcus aureus and Pseudomonas aeruginosa Biofilms.

Due to an increase in underlying predisposing factors, chronic wounds have become an increasing burden on healthcare systems worldwide. Chronic infections often contain biofilm-forming bacteria, which are challenging to eradicate due to increased antibiotic tolerance; thus, new and improved therapeutic strategies are warranted. One such strategy is the combination of ultrasound and antibiotics. Therefore, this study aimed to investigate the combinatory effects of low-frequency (50 kHz) ultrasound delivered by specially designed ultrasound patches using flexible piezoelectric material, PiezoPaint™, in combination with antibiotics against biofilms with Staphylococcus aureus and Pseudomonas aeruginosa. The reduction in viable cells in S. aureus and P. aeruginosa biofilms was evaluated post-treatment with fusidic acid, clindamycin, ciprofloxacin, and colistin in combination with ultrasound treatment. Two-hour ultrasound treatment significantly increased the bactericidal effect of all four antibiotics, resulting in a 96−98% and 90−93% reduction in P. aeruginosa and S. aureus, respectively. In addition, an additive effect was observed when extending treatment to 4 h, resulting in >99% and 95−97% reduction in P. aeruginosa and S. aureus, respectively. These results contrasted the lack of effect observed when treating filter-biofilms with antibiotics alone. The combined effect of ultrasound and antibiotic treatment resulted in a synergistic effect, reducing the viability of the clinically relevant pathogens S. aureus and P. aeruginosa. The modularity of the specially designed patches intended for topical treatment holds promising applications as a supplement in chronic wound therapy. Further studies are warranted with clinically isolated strains and other clinically relevant antibiotics before proceeding to studies where safety and applicability are investigated.

Common genetic variation in humans impacts in vitro susceptibility to SARS-CoV-2 infection.

The host response to SARS-CoV-2, the etiologic agent of the COVID-19 pandemic, demonstrates significant inter-individual variability. In addition to showing more disease in males, the elderly, and individuals with underlying comorbidities, SARS-CoV-2 can seemingly render healthy individuals with profound clinical complications. We hypothesize that, in addition to viral load and host antibody repertoire, host genetic variants also impact vulnerability to infection. Here we apply human induced pluripotent stem cell (hiPSC)-based models and CRISPR-engineering to explore the host genetics of SARS-CoV-2. We demonstrate that a single nucleotide polymorphism (rs4702), common in the population at large, and located in the 3'UTR of the protease FURIN, impacts alveolar and neuron infection by SARS-CoV-2 in vitro. Thus, we provide a proof-of-principle finding that common genetic variation can impact viral infection, and thus contribute to clinical heterogeneity in SARS-CoV-2. Ongoing genetic studies will help to better identify high-risk individuals, predict clinical complications, and facilitate the discovery of drugs that might treat disease.

Dual-frequency transducer for nonlinear contrast agent imaging.

Detection of high-order nonlinear components issued from microbubbles has emerged as a sensitive method for contrast agent imaging. Nevertheless, the detection of these high-frequency components, including the third, fourth, and fifth harmonics, remains challenging because of the lack of transducer sensitivity and bandwidth. In this context, we propose a new design of imaging transducer based on a simple fabrication process for high-frequency nonlinear imaging. The transducer is composed of two elements: the outer low-frequency (LF) element was centered at 4 MHz and used in transmit mode, whereas the inner high-frequency (HF) element centered at 14 MHz was used in receive mode. The center element was pad-printed using a lead zirconate titanate (PZT) paste. The outer element was molded using a commercial PZT, and curved porous unpoled PZT was used as backing. Each piezoelectric element was characterized to determine the electromechanical performance with thickness coupling factor around 45%. After the assembly of the two transducer elements, hydrophone measurements (electroacoustic responses and radiation patterns) were carried out and demonstrated a large bandwidth (70% at -3 dB) of the HF transducer. Finally, the transducer was evaluated for contrast agent imaging using contrast agent microbubbles. The results showed that harmonic components (up to the sixth harmonic) of the microbubbles were successfully detected. Moreover, images from a flow phantom were acquired and demonstrated the potential of the transducer for high-frequency nonlinear contrast imaging.

Non-planar pad-printed thick-film focused high-frequency ultrasonic transducers for imaging and therapeutic applications.

Pad-printed thick-film transducers have been shown to be an interesting alternative to lapped bulk piezoceramics, because the film is deposited with the required thickness, size, and geometry, thus avoiding any subsequent machining to achieve geometrical focusing. Their electromechanical properties are close to those of bulk ceramics with similar composition despite having a higher porosity. In this paper, padprinted high-frequency transducers based on a low-loss piezoceramic composition are designed and fabricated. High-porosity ceramic cylinders with a spherical top surface are used as the backing substrate. The transducers are characterized in view of imaging applications and their imaging capabilities are evaluated with phantoms containing spherical inclusions and in different biological tissues. In addition, the transducers are evaluated for their capability to produce high-acoustic intensities at frequencies around 20 MHz. High-intensity measurements, obtained with a calibrated hydrophone, show that transducer performance is promising for applications that would require the same device to be used for imaging and for therapy. Nevertheless, the transducer design can be improved, and simulation studies are performed to find a better compromise between low-power and high-power performance. The size, geometry, and constitutive materials of optimized configurations are proposed and their feasibility is discussed.

Fabrication of high-frequency pMUT arrays on silicon substrates.

A novel technique based on silicon micromachining for fabrication of linear arrays of high-frequency piezoelectric micromachined ultrasound transducers (pMUT) is presented. Piezoelectric elements are formed by deposition of lead zirconia titanate into etched features of a silicon substrate such that the depth of these features determine the element thickness and hence the resonance frequency. The process leaves a near planar surface which is ideal for further wafer level processing such as top electrode and interconnect formation. A fabricated element is characterized by pulse echo response.

3-D numerical modeling for axisymmetrical piezoelectric structures: application to high-frequency ultrasonic transducers.

The transient analysis of piezoelectric transducers is often performed using finite-element or finite-difference time-domain methods, which efficiently calculate the vibration of the structure but whose numerical dispersion prevents the modeling of waves propagating over large distances. A second analytical or numerical simulation is therefore often required to calculate the pressure field in the propagating medium (typically water) to deduce many important characteristics of the transducer, such as spatial resolutions and side lobe levels. This is why a hybrid algorithm was developed, combining finite- difference and pseudo-spectral methods in the case of 2-D configurations to simulate accurately both the generation of acoustic waves by the piezoelectric transducer and their propagation in the surrounding media using a single model. The algorithm was redefined in this study to take all three dimensions into account and to model single-element transducers, which usually present axisymmetrical geometry. This method was validated through comparison of its results with those of finite-element software, and was used to simulate the behavior of planar and lens-focused transducers. A high-frequency (30 MHz) transducer based on a screen-printed piezoelectric thick film was fabricated and characterized. The numerical results of the hybrid algorithm were found to be in good agreement with the experimental measurements of displacements at the surface of the transducer and of pressure radiated in water in front of the transducer.