Antimicrobial Effects of a Hexapetide KCM21 against Pseudomonas syringae pv. tomato DC3000 and Clavibacter michiganensis subsp. michiganensis

Antimicrobial peptides (AMPs) are small but effective cationic peptides with variable length. In previous study, four hexapeptides were identified that showed antimicrobial activities against various phytopathogenic bacteria. KCM21, the most effective antimicrobial peptide, was selected for further analysis to understand its modes of action by monitoring inhibitory effects of various cations, time-dependent antimicrobial kinetics, and observing cell disruption by electron microscopy. The effects of KCM21 on Gram-negative strain, Pseudomonas syringae pv. tomato DC3000 and Gram-positive strain, Clavibacter michiganensis subsp. michiganensis were compared. Treatment with divalent cations such as Ca²⁺ and Mg²⁺ inhibited the bactericidal activities of KCM21 significantly against P. syringae pv. tomato DC3000. The bactericidal kinetic study showed that KCM21 killed both bacteria rapidly and the process was faster against C. michiganensis subsp. michiganensis. The electron microscopic analysis revealed that KCM21 induced the formation of micelles and blebs on the surface of P. syringae pv. tomato DC3000 cells, while it caused cell rupture against C. michiganensis subsp. michiganensis cells. The outer membrane alteration and higher sensitivity to Ca²⁺ suggest that KCM21 interact with the outer membrane of P. syringae pv. tomato DC3000 cells during the process of killing, but not with C. michiganensis subsp. michiganensis cells that lack outer membrane. Considering that both strains had similar sensitivity to KCM21 in LB medium, outer membrane could not be the main target of KCM21, instead common compartments such as cytoplasmic membrane or internal macromolecules might be a possible target(s) of KCM21

Macrophage-Based Microrobots for Anticancer Therapy: Recent Progress and Future Perspectives

Macrophages, which are part of the mononuclear phagocytic system, possess sensory receptors that enable them to target cancer cells. In addition, they are able to engulf large amounts of particles through phagocytosis, suggesting a potential “Trojan horse” drug delivery approach to tumors by facilitating the engulfment of drug-hidden particles by macrophages. Recent research has focused on the development of macrophage-based microrobots for anticancer therapy, showing promising results and potential for clinical applications. In this review, we summarize the recent development of macrophage-based microrobot research for anticancer therapy. First, we discuss the types of macrophage cells used in the development of these microrobots, the common payloads they carry, and various targeting strategies utilized to guide the microrobots to cancer sites, such as biological, chemical, acoustic, and magnetic actuations. Subsequently, we analyze the applications of these microrobots in different cancer treatment modalities, including photothermal therapy, chemotherapy, immunotherapy, and various synergistic combination therapies. Finally, we present future outlooks for the development of macrophage-based microrobots

Two distinct actin waves correlated with turns-and-runs of crawling microglia

© 2019 Yang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.Freely crawling cells are often viewed as randomly moving Brownian particles but they generally exhibit some directional persistence. This property is often related to their zigzag motile behaviors that can be described as a noisy but temporally structured sequence of “runs” and “turns.” However, its underlying biophysical mechanism is largely unexplored. Here, we carefully investigate the collective actin wave dynamics associated with the zigzag-crawling movements of microglia (as primary brain immune cells) employing a number of different quantitative imaging modalities including synthetic aperture microscopy and optical diffraction tomography, as well as conventional fluorescence imaging and scanning electron microscopy. Interestingly, we find that microglia exhibit two distinct types of actin waves working at two quite different time scales and locations, and they seem to serve different purposes. One type of actin waves is fast “peripheral ruffles” arising spontaneously with an oscillating period of about 6 seconds at some portion of the leading edge of crawling microglia, where the vigorously biased peripheral ruffles seem to set the direction of a new turn (in 2-D free space). When the cell turning events are inhibited with a physical confinement (in 1-D track), the peripheral ruffles still exist at the leading edge with no bias but showing phase coherence in the cell crawling direction. The other type is “dorsal actin waves” which also exhibits an oscillatory behavior but with a much longer period of around 2 minutes compared to the fast “peripheral ruffles”. Dorsal actin waves (whether the cell turning events are inhibited or not) initiate in the lamellipodium just behind the leading edge, travelling down toward the core region of the cell and disappear. Such dorsal wave propagations seem to be correlated with migration of the cell. Thus, we may view the dorsal actin waves are connected with the “run” stage of cell body, whereas the fast ruffles at the leading front are involved in the “turn” stag

Four-Dimensional-Printed Microrobots and Their Applications: A Review

Owing to their small size, microrobots have many potential applications. In addition, four-dimensional (4D) printing facilitates reversible shape transformation over time or upon the application of stimuli. By combining the concept of microrobots and 4D printing, it may be possible to realize more sophisticated next-generation microrobot designs that can be actuated by applying various stimuli, and also demonstrates profound implications for various applications, including drug delivery, cells delivery, soft robotics, object release and others. Herein, recent advances in 4D-printed microrobots are reviewed, including strategies for facilitating shape transformations, diverse types of external stimuli, and medical and nonmedical applications of microrobots. Finally, to conclude the paper, the challenges and the prospects of 4D-printed microrobots are highlighted

Stable generation of multiple chemical gradients using in-situ formed nanoporous membranes

We developed a gradient generator of multiple chemicals using self-assembled particles in microchannels. The porous membranes based on the assembled particles allow the stable gradient generation without any convection flow disturbance and leakage. Moreover, they are more robust in dry environment and have long-term storage potential compared to hydrogel based membranes. Dynamic spatial control and the parallel preferential chemotaxis assay of salmonella typhimurium were successfully demonstrated. Our proposed devices are expected to be utilized for mimicking the real microenvironment for cells and various drug screening

Tryptophan-Rich and Proline-Rich Antimicrobial Peptides

Due to the increasing emergence of drug-resistant pathogenic microorganisms, there is a world-wide quest to develop new-generation antibiotics. Antimicrobial peptides (AMPs) are small peptides with a broad spectrum of antibiotic activities against bacteria, fungi, protozoa, viruses and sometimes exhibit cytotoxic activity toward cancer cells. As a part of the native host defense system, most AMPs target the membrane integrity of the microorganism, leading to cell death by lysis. These membrane lytic effects are often toxic to mammalian cells and restrict their systemic application. However, AMPs containing predominantly either tryptophan or proline can kill microorganisms by targeting intracellular pathways and are therefore a promising source of next-generation antibiotics. A minimum length of six amino acids is required for high antimicrobial activity in tryptophan-rich AMPs and the position of these residues also affects their antimicrobial activity. The aromatic side chain of tryptophan is able to rapidly form hydrogen bonds with membrane bilayer components. Proline-rich AMPs interact with the 70S ribosome and disrupt protein synthesis. In addition, they can also target the heat shock protein in target pathogens, and consequently lead to protein misfolding. In this review, we will focus on describing the structures, sources, and mechanisms of action of the aforementioned AMPs

Optimization of Field-Free Point Position, Gradient Field and Ferromagnetic Polymer Ratio for Enhanced Navigation of Magnetically Controlled Polymer-Based Microrobots in Blood Vessel

Microscale and nanoscale robots, frequently referred to as future cargo systems for targeted drug delivery, can effectively convert magnetic energy into locomotion. However, navigating and imaging them within a complex colloidal vascular system at a clinical scale is exigent. Hence, a more precise and enhanced hybrid control navigation and imaging system is necessary. Magnetic particle imaging (MPI) has been successfully applied to visualize the ensemble of superparamagnetic nanoparticles (MNPs) with high temporal sensitivity. MPI uses the concept of field-free point (FFP) mechanism in the principal magnetic field. The gradient magnetic field (|∇B|) of MPI scanners can generate sufficient magnetic force in MNPs; hence, it has been recently used to navigate nanosized particles and micron-sized swimmers. In this article, we present a simulation analysis of the optimized navigation of an ensemble of microsized polymer MNP-based drug carriers in blood vessels. Initially, an ideal two-dimensional FFP case is employed for the basic optimization of the FFP position to achieve efficient navigation. Thereafter, a nine-coil electromagnetic actuation simulation system is developed to generate and manipulate the FFP position and |∇B|. Under certain vessel and fluid conditions, the particle trajectories of different ferromagnetic polymer ratios and |∇B| were compared to optimize the FFP position

Concentration gradient generation of multiple chemicals using spatially controlled self-assembly of particles in microchannels

We present a robust microfluidic platform for the stable generation of multiple chemical gradients simultaneously using in situ self-assembly of particles in microchannels. This proposed device enables us to generate stable and reproducible diffusion-based gradients rapidly without convection flow: gradients are stabilized within 5 min and are maintained steady for several hours. Using this device, we demonstrate the dynamic position control of bacteria by introducing the sequential directional change of chemical gradients. Green Fluorescent Protein (GFP)-expressing bacterial cells, allowing quantitative monitoring, show not only tracking motion according to the directional control of chemical gradients, but also the gradual loss of sensitivity when exposed to the sequential attractants because of receptor saturation. In addition, the proposed system can be used to study the preferential chemotaxis assay of bacteria toward multiple chemical sources, since it is possible to produce multiple chemical gradients in the main chamber; aspartate induces the most preferential chemotaxis over galactose and ribose. The microfluidic device can be easily fabricated with a simple and cost effective process based on capillary pressure and evaporation for particle assembly. The assembled particles create uniform porous membranes in microchannels and its porosity can be easily controlled with different size particles. Moreover, the membrane is biocompatible and more robust than hydrogel-based porous membranes. The proposed system is expected to be a useful tool for the characterization of bacterial responses to various chemical sources, screening of bacterial cells, synthetic biology and understanding many cellular activities.close10

Acoustic Actuators for the Manipulation of Micro/Nanorobots: State-of-the-Art and Future Outlooks

Compared to other actuating methods, acoustic actuators offer the distinctive capability of the contactless manipulation of small objects, such as microscale and nanoscale robots. Furthermore, they have the ability to penetrate the skin, allowing for the trapping and manipulation of micro/nanorobots that carry therapeutic agents in diverse media. In this review, we summarize the current progress in using acoustic actuators for the manipulation of micro/nanorobots used in various biomedical applications. First, we introduce the actuating method of using acoustic waves to manipulate objects, including the principle of operation and different types of acoustic actuators that are usually employed. Then, applications involving manipulating different types of devices are reviewed, including bubble-based microrobots, bubble-free robots, biohybrid microrobots, and nanorobots. Finally, we discuss the challenges and future perspectives for the development of the field