Applications of upconversion nanoparticles in cellular optogenetics.

Upconversion-mediated optogenetics is an emerging powerful technique to remotely control and manipulate the deep-tissue protein functions and signaling pathway activation. This technique uses lanthanide upconversion nanoparticles (UCNPs) as light transducers and through near-infrared light to indirectly activate the traditional optogenetic proteins. With the merits of high spatiotemporal resolution and minimal invasiveness, this technique enables cell-type specific manipulation of cellular activities in deep tissues as well as in living animals. In this review, we introduce the latest development of optogenetic modules and UCNPs, with emphasis on the integration of UCNPs with cellular optogenetics and their biomedical applications on the control of neural/brain activity, cancer therapy and cardiac optogenetics in vivo. Furthermore, we analyze the current developed strategies to optimize and advance the upconversion-mediated optogenetics and discuss the remaining challenges of its further applications in biomedical study and clinical translational research. STATEMENT OF SIGNIFICANCE: Optogenetics harnesses photoactivatable proteins to optically stimulate and control intracellular activities. UCNPs-mediated NIR-activatable optogenetics uses lanthanide upconversion nanoparticles (UCNPs) as light transducers and utilizes near-infrared (NIR) light to indirectly activate the traditional optogenetic proteins. The integration of UCNPs with cellular optogenetics has showed great promise in biomedical applications in regulating neural/brain activity, cancer therapy and cardiac optogenetics in vivo. The evolution and optimization of functional UCNPs and the discovery and engineering of novel optogenetic modules would both contribute to the advance of such unique hybrid technology, which may lead to discoveries in biomedical research and provide new treatments for human diseases.

Structural insights into phospholipase D function.

Phospholipase D (PLD) and its metabolic active product phosphatidic acid (PA) engage in a wide range of physiopathologic processes in the cell. PLDs have been considered as a potential and promising drug target. Recently, the crystal structures of PLDs in mammalian and plant have been solved at atomic resolution. These achievements allow us to understand the structural differences among different species of PLDs and the functions of their key domains. In this review, we summarize the sequence and structure of different species of PLD isoforms, and discuss the structural mechanisms for PLD interactions with their binding partners and the functions of each key domain in the regulation of PLDs activation and catalytic reaction.

Programmable Transformation and Controllable Locomotion of Magnetoactive Soft Materials with 3D-Patterned Magnetization.

Magnetoactive soft material (MASM) is distinguished for multifunctional shape manipulations under magnetic actuation, thereby holding a great promise in soft robotics, actuators, electronics, and metamaterials. However, the current research of MASM with continuum hard-magnetic profiles focuses little on the transformation mechanism, high dimensional shape transformation, and multistable locomotion. Herein, we developed a systematic methodology for programmable transformation and controllable locomotion of MASM with 3D-patterned continuum magnetization. An iterative computational model based on the equilibrium between magnetic torque and deformation-induced elastic torque was developed for precise prediction of MASM transformation. Multidimensional and complex shape manipulation ability of MASM was demonstrated by magnetically actuated transformations, including 1D to 2D, 2D to 3D, and 3D to 4D transformations of solid MASM, 2D to 3D pattern transformation of MASM-based elastin-like mesh, and 3D to 4D transformation of MASM-based cuboidal lattice. Multistable and controllable locomotion of MASM was verified by multimodal locomotion behaviors of a scallop-inspired robot for wall climbing in a dry frame and drug delivery in wet stomach, including roll, open, and close under self-locked and unlocked states.

Dual Function of PI(4,5)P2 in Insulin-Regulated Exocytic Trafficking of GLUT4 in Adipocytes.

Phosphoinositides are important signaling molecules involved in the regulation of vesicular trafficking. It has been implicated that phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] is involved in insulin-regulated GLUT4 translocation in adipocytes. However, it remains unclear where and how PI(4,5)P2 regulates discrete steps of GLUT4 vesicle translocation in adipocytes, especially on the exocytic arm of regulation. Here, we employed optogenetic tools to acutely control the PI(4,5)P2 metabolism in living cells. By combination of TIRFM imaging, we were able to monitor the temporal-spatial-dependent PI(4,5)P2 regulation on discrete steps of GLUT4 translocation in adipocytes. We found that the plasma membrane localized PI(4,5)P2 is crucial for proper insulin signaling propagation and for insulin-stimulated GLUT4 vesicle translocation in 3T3-L1 adipocytes. Global depletion of PI(4,5)P2 on the cell surface blunted insulin-stimulated Akt phosphorylation and abolished insulin effects in promotion of the docking and fusion of GLUT4 vesicle with the plasma membrane. Furthermore, by development of a novel optogenetic module to selectively modulate PI(4,5)P2 levels on the GLUT4 vesicle docking site, we identified an important regulatory role of PI(4,5)P2 in controlling of vesicle docking process. Local depletion of PI(4,5)P2 at the vesicle docking site promoted GLUT4 vesicle undocking, diminished insulin-stimulated GLUT4 vesicle docking and fusion, but without perturbation of insulin signaling propagation in adipocytes. Our results provide strong evidence that cell surface PI(4,5)P2 plays two distinct functions on regulation of the exocytic trafficking of GLUT4 in adipocytes. PI(4,5)P2 not only regulates the proper activation of insulin signaling in general but also controls GLUT4 vesicle docking process at the vesicle-membrane contact sites.

Rapidly Fabricated Microneedle Arrays Using Magnetorheological Drawing Lithography for Transdermal Drug Delivery.

Microneedle arrays (MAs) are among the most promising transdermal drug delivery systems in the last decades due to its minimally invasive nature, convenient operation, and first-pass-metabolism avoidance. However, most MA fabrication methods are difficult to operate, need multiple steps, or require expensive equipment. A novel magnetorheological drawing lithography approach was proposed to rapidly fabricate a flexible microneedle array (FMA) for transdermal drug delivery. A 3D structural liquid MA was drawn in one step from the droplets of curable magnetorheological fluid and maintained its shape under an external magnetic field. The liquid MA was subsequently solidified and sputter-coated with the Ti/Au film to fabricate FMA. FMA morphology, mechanical properties, and transdermal drug delivery performance in vitro were experimentally investigated. FMA consisted of a 5 × 5 cone-shaped microneedle array on a PET flexible substrate. FMA exhibited good strength and excellent penetration performance. It could easily penetrate into skin without breakage, creating microchannels for the promotion of skin permeability. Drugs could be well permeated and diffused in the skin along the microchannels created by FMA. Finally, a dissolvable microneedle array (DMA) was also fabricated by a micromolding technique using FMA as a master template. The DMA exhibited good dissolvable and permeable performance in the agarose block.

Touch-actuated microneedle array patch for closed-loop transdermal drug delivery.

To date, only approximately 20 drugs synthesized with small molecules have been approved by the FDA for use in traditional transdermal patches (TTP) owing to the extremely low permeation rate of the skin barrier for macromolecular drugs. A novel touch-actuated microneedle array patch (TMAP) was developed for transdermal delivery of liquid macromolecular drugs. TMAP is a combination of a typical TTP and a solid microneedle array (MA). High doses of liquid drug formulations, especially heat-sensitive compounds can be easily filled and stored in the drug reservoir of TMAPs. TMAP can easily penetrate the skin and automatically retract from it to create microchannels through the stratum corneum (SC) layer using touch-actuated 'press and release' actions for passive permeation of liquid drugs. Comparison of subcutaneous injection, TTP, solid MA, and dissolvable MA, indicated that insulin-loaded TMAP exhibited the best hypoglycemic effect on type 1 diabetic rats. A 'closed-loop' permeation control was also provided for on-demand insulin delivery based on feedback of blood glucose levels (BGLs). Twenty IU-insulin-loaded TMAP maintained the type 1 diabetic rats in a normoglycemic state for approximately 11.63 h, the longest therapeutic duration among all previously reported results on microneedle-based transdermal patches. TMAP possesses excellent transdermal drug delivery capabilities.

Additive Manufacturing of Honeybee-Inspired Microneedle for Easy Skin Insertion and Difficult Removal.

With natural evolution, honeybee stinger with microbarbs can easily penetrate and trap in the skin of hostile animals to inject venom for self-defense. We proposed a novel three-dimensional additive manufacturing method, namely magnetorheological drawing lithography, to efficiently fabricate a bioinspired microneedle imitating a honeybee stinger. Under the assistance of an external magnetic field, a parent microneedle was directly drawn on the pillar tip, and tilted microbarbs were subsequently formed on the four sides of the parent microneedle. Compared with the barbless microneedle, the microstructured barbs enable the bioinspired microneedle for easy skin insertion and difficult removal. The extraction-penetration force ratio of the bioinspired microneedle was triple that of the barbless microneedle. The stress concentration at the barbs helps to reduce the insertion force of the bioinspired microneedle by minimizing the frictional force, whereas it increases the adhesion force by interlocking the barbs in the tissue during retraction. Such finds may provide an inspiration for further design of barbed microtip-based microneedles for tissue adhesion, transdermal drug delivery, biosignal recording, and so on.

Sleep Apnea Detection Based on Thoracic and Abdominal Movement Signals of Wearable Piezo-Electric Bands.

Physiologically, the thoracic (THO) and abdominal (ABD) movement signals, captured using wearable piezo-electric bands, provide information about various types of apnea, including central sleep apnea (CSA) and obstructive sleep apnea (OSA). However, the use of piezo-electric wearables in detecting sleep apnea events has been seldom explored in the literature. This study explored the possibility of identifying sleep apnea events, including OSA and CSA, by solely analyzing one or both the THO and ABD signals. An adaptive non-harmonic model was introduced to model the THO and ABD signals, which allows us to design features for sleep apnea events. To confirm the suitability of the extracted features, a support vector machine was applied to classify three categories - normal and hypopnea, OSA, and CSA. According to a database of 34 subjects, the overall classification accuracies were on average 75.9%±11.7% and 73.8%±4.4%, respectively, based on the cross validation. When the features determined from the THO and ABD signals were combined, the overall classification accuracy became 81.8%±9.4%. These features were applied for designing a state machine for online apnea event detection. Two event-byevent accuracy indices, S and I, were proposed for evaluating the performance of the state machine. For the same database, the S index was 84.01%±9.06%, and the I index was 77.21%±19.01%. The results indicate the considerable potential of applying the proposed algorithm to clinical examinations for both screening and homecare purposes.