Direct Reprogramming of Rat Neural Precursor Cells and Fibroblasts into Pluripotent Stem Cells

Background: Given the usefulness of rats as an experimental system, an efficient method for generating rat induced pluripotent stem (iPS) cells would provide researchers with a powerful tool for studying human physiology and disease. Here, we report direct reprogramming of rat neural precursor (NP) cells and rat embryonic fibroblasts (REF) into iPS cells by retroviral transduction using either three (Oct3/4, Sox2, and Klf4), four (Oct3/4, Sox2, Klf4, and c-Myc), or five (Oct3/4, Sox2, Klf4, c-Myc, and Nanog) genes. Methodology and Principal Findings: iPS cells were generated from both NP and REF using only three (Oct3/4, Sox2, and Klf4) genes without c-Myc. Two factors were found to be critical for efficient derivation and maintenance of rat iPS cells: the use of rat instead of mouse feeders, and the use of small molecules specifically inhibiting mitogen-activated protein kinase and glycogen synthase kinase 3 pathways. In contrast, introduction of embryonic stem cell (ESC) extracts induced partial reprogramming, but failed to generate iPS cells. However, when combined with retroviral transduction, this method generated iPS cells with significantly higher efficiency. Morphology, gene expression, and epigenetic status confirmed that these rat iPS cells exhibited ESC-like properties, including the ability to differentiate into all three germ layers both in vitro and in teratomas. In particular, we found that these rat iPS cells could differentiate to midbrain-like dopamine neurons with a high efficiency. Conclusions/Significance: Given the usefulness of rats as an experimental system, our optimized method would be useful for generating rat iPS cells from diverse tissues and provide researchers with a powerful tool for studying human physiology and disease

Spatio-spectral 4D coherent ranging using a flutter-wavelength-swept laser

Abstract Coherent light detection and ranging (LiDAR), particularly the frequency-modulated continuous-wave LiDAR, is a robust optical imaging technology for measuring long-range distance and velocity in three dimensions (3D). We propose a spatio-spectral coherent LiDAR based on a unique wavelength-swept laser to enable both axial coherent ranging and lateral spatio-spectral beam scanning simultaneously. Instead of the conventional unidirectional wavelength-swept laser, a flutter-wavelength-swept laser (FWSL) successfully decoupled bidirectional wavelength modulation and continuous wavelength sweep, which overcame the measurable distance limited by the sampling process. The decoupled operation in FWSL enabled sequential sampling of flutter-wavelength modulation across its wide spectral bandwidth of 160 nm and, thus, allowed simultaneous distance and velocity measurement over an extended measurable distance. Herein, complete four-dimensional (4D) imaging, combining real-time 3D distance and velocity measurements, was implemented by solid-state beam scanning. An acousto-optic scanner was synchronized to facilitate the other lateral beam scanning, resulting in an optimized solid-state coherent LiDAR system. The proposed spatio-spectral coherent LiDAR system achieved high-resolution coherent ranging over long distances and real-time 4D imaging with a frame rate of 10 Hz, even in challenging environments

Micropatterning of metal oxide nanofibers by electrohydrodynamic (EHD) printing towards highly integrated and multiplexed gas sensor applications

Integration of heterogeneous sensing materials in microelectronic devices is essential to accomplish compact and highly integrated environmental sensors. For this purpose, a micro-patterning method of electrospun metal oxide nanofibers based on electrohydrodynamic (EHD) printing process was developed in this work. Several types of metal oxide (SnO_2, In_2O_3, WO_3 and NiO) nanofibers that were produced by electrospinning, fragmented into smaller pieces by ultrasonication, and dissolved in organic solvents were utilized as inks for the printing. Constant or pulsed wave bias consisting of base and jetting voltages were applied between a nozzle and a substrate to generate a jetting of nanofiber solutions. Several parameters for EHD printing such as pulse width, inner diameter of the nozzle, distance from the nozzle to the substrate, and stage speed, were optimized for accurate micro-patterning of electrospun nanofibers. By using optimized printing parameters, microscale patterns of electrospun nanofibers with a minimum diameter less than 50 μm could be realized. Gas sensors were fabricated by EHD printing on the microelectrodes and then used for the detection of toxic gases such as NO_2, CO and H_2S. Four kinds of metal oxides could detect down to 0.1 ppm of NO_2, 1 ppm of H_2S and 20 ppm of CO gases. Also, heterogeneous nanofiber gas sensor array was fabricated by the same printing method and could detect NO_2 using the sensor array platform with microheaters. Furthermore, microscale patterns of nanofibers by EHD printing could be applied to the suspended MEMS platform without any structural damage and this sensor array could detect NO_2 and H_2S gases with 20 mW power consumption

Electrochemical Generation of Mesopores and Residual Oxygen for the Enhanced Activity of Silver Electrocatalysts

The development of stable and efficient electrocatalysts is of key importance for the establishment of a sustainable society. The activity of a metal electrocatalyst is determined by its electrochemically active surface area and intrinsic activity, which can be increased using highly porous structures and heteroatomic doping, respectively. Herein, we propose a general strategy of generating mesopores and residual oxygen in metal electrocatalysts by reduction of metastable metal oxides using Ag2O3 electrodeposited onto carbon paper as a model system and demonstrating that the obtained multipurpose porous Ag electrocatalyst has high activity for the electroreduction of O-2 and CO2. The presence of mesopores and residual oxygen is confirmed by electrochemical and spectroscopic techniques, and quantum mechanical simulations prove the importance of residual oxygen for electrocatalytic activity enhancement. Thus, the adopted strategy is concluded to allow the synthesis of highly active metal catalysts with controlled mesoporosity and residual oxygen content

Photoswitchable Microgels for Dynamic Macrophage Modulation

Dynamic manipulation of supramolecular self-assembled structures is achieved irreversibly or under non-physiological conditions, thereby limiting their biomedical, environmental, and catalysis applicability. In this study, microgels composed of azobenzene derivatives stacked via pi-cation and pi-pi interactions are developed that are electrostatically stabilized with Arg-Gly-Asp (RGD)-bearing anionic polymers. Lateral swelling of RGD-bearing microgels occurs via cis-azobenzene formation mediated by near-infrared-light-upconverted ultraviolet light, which disrupts intermolecular interactions on the visible-light-absorbing upconversion-nanoparticle-coated materials. Real-time imaging and molecular dynamics simulations demonstrate the deswelling of RGD-bearing microgels via visible-light-mediated trans-azobenzene formation. Near-infrared light can induce in situ swelling of RGD-bearing microgels to increase RGD availability and trigger release of loaded interleukin-4, which facilitates the adhesion structure assembly linked with pro-regenerative polarization of host macrophages. In contrast, visible light can induce deswelling of RGD-bearing microgels to decrease RGD availability that suppresses macrophage adhesion that yields pro-inflammatory polarization. These microgels exhibit high stability and non-toxicity. Versatile use of ligands and protein delivery can offer cytocompatible and photoswitchable manipulability of diverse host cells