Hub Proteins Involved in RAW 264.7 Macrophages Exposed to Direct Current Electric Field.

At present, studies on macrophage proteins mainly focus on biological stimuli, with less attention paid to the responses of macrophage proteins to physical stimuli, such as electric fields. Here, we exploited the electric field-sensitive hub proteins of macrophages. RAW 264.7 macrophages were treated with a direct current electric field (dcEF) (200 mV/mm) for four hours, followed by RNA-Seq analysis. Differentially expressed genes (DEGs) were obtained, followed by Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes pathway (KEGG) and protein-protein interaction (PPI) analysis. Eight qPCR-verified DEGs were selected. Subsequently, three-dimensional protein models of DEGs were modeled by Modeller and Rosetta, followed by molecular dynamics simulation for 200 ns with GROMACS. Finally, dcEFs (10, 50, and 500 mV/mm) were used to simulate the molecular dynamics of DEG proteins for 200 ns, followed by trajectory analysis. The dcEF has no obvious effect on RAW 264.7 morphology. A total of 689 DEGs were obtained, and enrichment analysis showed that the steroid biosynthesis pathway was most affected by the dcEF. Moreover, the three-dimensional protein structures of hub proteins were constructed, and trajectory analysis suggested that the dcEF caused an increase in the atomic motion of the protein in a dcEF-intensity-dependent manner. Overall, we provide new clues and a basis for investigating the hub proteins of macrophages in response to electric field stimulation.

Cellular Processes Involved in Jurkat Cells Exposed to Nanosecond Pulsed Electric Field.

Recently, nanosecond pulsed electric field (nsPEF) has been considered as a new tool for tumor therapy, but its molecular mechanism of function remains to be fully elucidated. Here, we explored the cellular processes of Jurkat cells exposed to nanosecond pulsed electric field. Differentially expressed genes (DEGs) were acquired from the GEO2R, followed by analysis with a series of bioinformatics tools. Subsequently, 3D protein models of hub genes were modeled by Modeller 9.21 and Rosetta 3.9. Then, a 100 ns molecular dynamics simulation for each hub protein was performed with GROMACS 2018.2. Finally, three kinds of nsPEF voltages (0.01, 0.05, and 0.5 mV/mm) were used to simulate the molecular dynamics of hub proteins for 100 ns. A total of 1769 DEGs and eight hub genes were obtained. Molecular dynamic analysis, including root mean square deviation (RMSD), root mean square fluctuation (RMSF), and the Rg, demonstrated that the 3D structure of hub proteins was built, and the structural characteristics of hub proteins under different nsPEFs were acquired. In conclusion, we explored the effect of nsPEF on Jurkat cell signaling pathway from the perspective of molecular informatics, which will be helpful in understanding the complex effects of nsPEF on acute T-cell leukemia Jurkat cells.

Light addressable potentiometric sensor with well-ordered pyramidal pits-patterned silicon.

Light addressable potentiometric sensor (LAPS) with the structure of electrolyte-insulator-semiconductor is a kind of field effect sensor that detects local potential changes caused by protonation and deprotonation between electrolyte and insulator by light pulse. And scanned light pulse allows two-dimensional imaging of the distribution of chemical/biological species on the surface of sensor. An important challenge is to achieve low-cost, strong anti-interference and high-performance silicon-based LAPS. In this study, we propose to combine microsphere lithography with wet etching to fabricate well-ordered, tunable and low-cost pyramidal pits-patterned silicon as semiconductor of LAPS. The morphology and optical properties of pyramidal pits-patterned silicon are tested and analyzed. The sensing characteristics and the pH imaging performance of LAPS are tested and evaluated. The experiment results and theoretical analyses show that LAPS with pyramidal pits-patterned silicon has acceptable pH response, good long-term stability, and high performance in terms of photocurrent enhancement ratio, signal-to-noise ratio and pH imaging. This work can provide a simple, low-cost, strong anti-interference and high-performance device for pH-related chemical/biological analysis.

Light Addressable Potentiometric Sensors for Biochemical Imaging on Microscale: A Review on Optimization of Imaging Speed and Spatial Resolution.

Light addressable potentiometric sensors (LAPS) are a competitive tool for unmarked biochemical imaging, especially imaging on microscale. It is essential to optimize the imaging speed and spatial resolution of LAPS since the imaging targets of LAPS, such as cell, microfluidic channel, etc., require LAPS to image at the micrometer level, and a fast enough imaging speed is a prerequisite for the dynamic process involved in biochemical imaging. In this study, we discuss the improvement of LAPS in terms of imaging speed and spatial resolution. The development of LAPS in imaging speed and spatial resolution is demonstrated by the latest applications of biochemistry monitoring and imaging on the microscale.

Honeycomb meshed working electrodes based on microsphere lithography for high-resolution chemical image sensor.

The chemical image sensor has attracted much attention because of its ability to visualize the spatial distribution of biochemical species in solution. However, the lateral diffusion of photo-generated carriers generated by illuminating backside of the substrate limits the spatial resolution of the sensor and the sharpness of the image. In this study, honeycomb meshed metal array based on microsphere lithography as working electrodes of the sensor is proposed. A focused light spot illuminates a plurality of periodic and ordered mesh holes and the metal electrodes adjacent to the mesh holes. For chemical imaging performance, sensors with honeycomb meshed working electrodes are developed for comparison with the sensor the flat working electrodes, and their spatial resolution is tested under the focused light spot with different sizes. Three interpretations of improved spatial resolution including enhanced recombination of the photo-generated holes by electrons during the initial stage of lateral diffusion and two boundary cases of modulated light illuminating meshed working electrodes on the substrate are elaborated. The experimental results show that the spatial resolution of sensor chips with honeycomb meshed working electrodes is better than that of the sensor with the flat working electrodes and is higher under the illumination condition of smaller light spots.

Cellular processes involved in lung cancer cells exposed to direct current electric field.

With the rapid breakthrough of electrochemical treatment of tumors, electric field (EF)-sensitive genes, previously rarely exploited, have become an emerging field recently. Here, we reported our work for the identification of EF-sensitive genes in lung cancer cells. The gene expression profile (GSE33845), in which the human lung cancer CL1-0 cells were treated with a direct current electric field (dcEF) (300 mV/mm) for 2 h, was retrieved from GEO database. Differentially expressed genes (DEGs) were acquired, followed by Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes pathway (KEGG) and protein-protein interaction (PPI) analysis. Hub genes were acquired and analyzed by various tools including the Human Protein Atlas, Kaplan-Meier analysis, Cytoscape, FunRich, Oncomine and cBioPortal. Subsequently, three-dimensional protein models of hub genes were modeled by Modeller 9.20 and Rosetta 3.9. Finally, a 100 ns molecular dynamics simulation for each hub protein was performed with GROMACS 2018.2. A total of 257 DEGs were acquired and analyzed by GO, KEGG and PPI. Then, 10 hub genes were obtained, and the signal pathway analysis showed that two inflammatory pathways were activated: the FoxO signaling pathway and the AGE-RAGE signaling pathway. The molecular dynamic analysis including RMSD and the radius of gyration hinted that the 3D structures of hub proteins were built. Overall, our work identified EF-sensitive genes in lung cancer cells and identified that the inflammatory state of tumor cells may be involved in the feedback mechanism of lung cancer cells in response to electric field stimulation. In addition, qualified three-dimensional protein models of hub genes were also constructed, which will be helpful in understanding the complex effects of dcEF on human lung cancer CL1-0 cells.