CRISPR/Cas-based Diagnostics and Gene Therapy

Clustered regularly interspaced short palindromic repeats (CRISPR) technology, an easy, rapid, cost-effective, and precise gene-editing technique, has revolutionized diagnostics and gene therapy. Fast and accurate diagnosis of diseases is essential for point-of-care-testing (POCT) and specialized medical institutes. The CRISPR-associated (Cas) proteins system shed light on the new diagnostics methods at point-of-care (POC) owning to its advantages. In addition, CRISPR/Cas-based gene-editing technology has led to various breakthroughs in gene therapy. It has been employed in clinical trials for a variety of untreatable diseases, including cancer, blood disorders, and other syndromes. Currently, the clinical application of CRISPR/Cas has been mainly focused on ex vivo therapies. Recently, tremendous efforts have been made in the development of ex vivo gene therapy based on CRISPR-Cas9. Despite these efforts, in vivo CRISPR/Cas gene therapy is only in its initial stage. Here, we review the milestones of CRISPR/Cas technologies that advanced the field of diagnostics and gene therapy. We also highlight the recent advances of diagnostics and gene therapy based on CRISPR/Cas technology. In the last section, we discuss the strength and significant challenges of the CRISPR/Cas technology for its future clinical usage in diagnosis and gene therapy.

Synergistic Effects of Acoustics-based Therapy and Immunotherapy in Cancer Treatment

Cancer is an intractable disease and has ability to escape immunological recognition. Cancer immunotherapy to enhance the autogenous immune response to cancer tissue is reported to be the most promising method for cancer treatment. After the release of damage-associated molecular patterns, dendritic cells come mature and then recruit activated T cells to induce immune response. To trigger the release of cancer associated antigens, cancer acoustics-based therapy has various prominent advantages and has been reported in various research. In this review, we classified the acoustics-based therapy into sonopyrolysis-, sonoporation-, and sonoluminescence-based therapy. Then, detailed mechanisms of these therapies are discussed to show the status of cancer immunotherapy induced by acoustics-based therapy in quo. Finally, we express some future prospects in this research field and make some predictions of its development direction

Progress in the Application of Nano- and Micro-based Drug Delivery Systems in Pulmonary Drug Delivery

Nanotechnology is associated with the development of particles in the nano-size range that can be used in a wide range of applications in the medical field. It has gained more importance in the pharmaceutical research field particularly in drug delivery, as it results in enhanced therapeutic drug performance, improved drug solubility, targeted drug delivery to the specific sites, minimized side effects, and prolonged drug retention time in the targeted site. To date, the application of nanotechnology continues to offer several benefits in the treatment of various chronic diseases and results in remarkable improvements in treatment outcomes. The use of nano-based delivery systems such as liposomes, micelles, and nanoparticles in pulmonary drug delivery have shown to be a promising strategy in achieving drug deposition and maintained controlled drug release in the lungs. They have been widely used to minimize the risks of drug toxicity in vivo. In this review, recent advances in the application of nano- and micro-based delivery systems in pulmonary drug delivery for the treatment of various pulmonary diseases, such as lung cancer, asthma, and chronic obstructive pulmonary disease, are highlighted. Limitations in the application of these drug delivery systems and some key strategies in improving their formulation properties to overcome challenges encountered in drug delivery are also discussed.

Sonoporation: Underlying Mechanisms and Applications in Cellular Regulation

Ultrasound combined with microbubble-mediated sonoporation has been applied to enhance drug or gene intracellular delivery. Sonoporation leads to the formation of openings in the cell membrane, triggered by ultrasound-mediated oscillations and destruction of microbubbles. Multiple mechanisms are involved in the occurrence of sonoporation, including ultrasonic parameters, microbubbles size, and the distance of microbubbles to cells. Recent advances are beginning to extend applications through the assistance of contrast agents, which allow ultrasound to connect directly to cellular functions such as gene expression, cellular apoptosis, differentiation, and even epigenetic reprogramming. In this review, we summarize the current state of the art concerning microbubble–cell interactions and sonoporation effects leading to cellular functions.

Nanoparticles Enable Efficient Delivery of Antimicrobial Peptides for the Treatment of Deep Infections

Antimicrobial peptides (AMPs) have emerged as promising alternatives of traditional antibiotics against drug-resistant bacteria owing to their broad-spectrum antimicrobial properties and low tendency to drugresistance. However, their therapeutic efficacy in vivo, especially for infections in deep organs, is limited owing to their systemic toxicity and low bioavailability. Nanoparticles-based delivery systems offer a strategy to increase the therapeutic index of AMPs by preventing proteolysis, increasing the accumulation at infection sites, and reducing toxicity. Herein, we will discuss the current progress of using nanoparticles as delivery vehicles for AMPs for the treatment of deep infections.

Deep impact of superficial skin inking: acoustic analysis of underlying tissue

Background: Skin tattoos are a common decoration, but profound scientific study whether the presence of a skin tattoo alters the acoustic response from superficial tissue, and therefore from underlying tissue, was previously lacking. Any image aberrations caused by tattoo presence may have been thought negligible, yet empirically found artifacts in brightness-mode images of tattooed skin suggest otherwise. This study investigated the nature of these artifacts theoretically and experimentally in extremely simplified cases of perfectly flat and homogenous layered media and in tattooed pork.Methods: Theory was derived for computing the acoustic response from horizontally and vertically layered media containing a thin inked layer. Experiments were performed in vitro. Artificial and pork skin were tattooed, attached to phantom material, and sonicated with a 13–6-MHz probe. The speed of sound of these materials was determined, and the perceived refraction angles was measured.Results: The measured speeds of sound of tattooed materials were higher than those of their uninked counterparts. The presence of tattoo ink was found to have increased the linear acoustic attenuation by 1 dB/cm. This value is negligible for typical tattoos of only few millimeters. The perceived critical refraction angles of adjacent materials could be detected, and their corresponding speeds of sound were quantified. These coincided with values derived from theory.Conclusion: The ratio of speeds of sound of adjacent materials was shown to create distinct highlights in brightness-mode images. The artifacts observed in in vitro and in vivo brightness-mode scans were explained from near-vertical transitions between areas of different sound speed. This is the first study correlating so-called critical refraction highlighting with speed-of-sound information. In addition, it was found that phantom material is a room-temperature acoustic alternative for experiments on live human skin. In summary, the presence of superficial tattoos has a small but quantifiable effect on the acoustic response from deeper tissues.