Lentviral-mediated RNAi to inhibit target gene expression of the porcine integrin αv subunit, the FMDV receptor, and against FMDV infection in PK-15 cells

<p>Abstract</p> <p>Background</p> <p>shRNA targeting the integrin αv subunit, which is the foot-and-mouth disease virus (FMDV) receptor, plays a key role in virus attachment to susceptible cells. We constructed a RNAi lentiviral vector, iαv pLenti6/BLOCK -iT™, which expressed siRNA targeting the FMDV receptor, the porcine integrin αv subunit, on PK-15 cells. We also produced a lentiviral stock, established an iαv-PK-15 cell line, evaluated the gene silencing efficiency of mRNA using real-time qRT-PCR, integrand αv expression by indirect immunofluorescence assay (IIF) and cell enzyme linked immunosorbent assays (cell ELISA), and investigated the in vivo inhibitory effect of shRNA on FMDV replication in PK-15 cells.</p> <p>Results</p> <p>Our results indicated successful establishment of the iαv U6 RNAi entry vector and the iαv pLenti6/BLOCK -iT expression vector. The functional titer of obtained virus was 1.0 × 10⁶TU/mL. To compare with the control and mock group, the iαv-PK-15 group αv mRNA expression rate in group was reduced by 89.5%, whilst IIF and cell ELISA clearly indicated suppression in the experimental group. Thus, iαv-PK-15 cells could reduce virus growth by more than three-fold and there was a > 99% reduction in virus titer when cells were challenged with 10²TCID₅₀of FMDV.</p> <p>Conclusions</p> <p>Iαv-PK-15 cells were demonstrated as a cell model for anti-FMDV potency testing, and this study suggests that shRNA could be a viable therapeutic approach for controlling the severity of FMD infection and spread.</p

Crosstalk Between Autophagy and Cerebral Ischemia

With the use of advanced electron microscopy and molecular biology tools, several studies have shown that autophagy is involved in the development of ischemic stroke. A series of molecular mechanisms are involved in the regulation of autophagy. In this work, the possible molecular mechanisms involved in autophagy during ischemic stroke were reviewed and new potential targets for the study and treatment of ischemic stroke were provided

The Advantage of Low-Delta Electroencephalogram Phase Feature for Reconstructing the Center-Out Reaching Hand Movements

It is an emerging frontier of research on the use of neural signals for prosthesis control, in order to restore lost function to amputees and patients after spinal cord injury. Compared to the invasive neural signal based brain-machine interface (BMI), a non-invasive alternative, i.e., the electroencephalogram (EEG)-based BMI would be more widely accepted by the patients above. Ideally, a real-time continuous neuroprosthestic control is required for practical applications. However, conventional EEG-based BMIs mainly deal with the discrete brain activity classification. Until recently, the literature has reported several attempts for achieving the real-time continuous control by reconstructing the continuous movement parameters (e.g., speed, position, etc.) from the EEG recordings, and the low-frequency band EEG is consistently reported to encode the continuous motor control information. Previous studies with executed movement tasks have extensively relied on the amplitude representation of such slow oscillations of EEG signals for building models to decode kinematic parameters. Inspired by the recent successes of instantaneous phase of low-frequency invasive brain signals in the motor control and sensory processing domains, this study examines the extension of such a slow-oscillation phase representation to the reconstructing two-dimensional hand movements, with the non-invasive EEG signals for the first time. The data for analysis are collected on five healthy subjects performing 2D hand center-out reaching along four directions in two sessions. On representative channels over the cortices encoding the execution information of reaching movements, we show that the low-delta EEG phase representation is characterized by higher signal-to-noise ratio and stronger modulation by the movement tasks, compared to the low-delta EEG amplitude representation. Furthermore, we have tested the low-delta EEG phase representation with two commonly used linear decoding models. The results demonstrate that the low-delta EEG phase based decoders lead to superior performance for 2D executed movement reconstruction to its amplitude based counterparts, as well as the other-frequency band amplitude and power based features. Thus, our study contributes to improve the movement reconstruction from EEG by introducing a new feature set based on the low-delta EEG phase patterns, and demonstrates its potential for continuous fine motion control of neuroprostheses

Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals &lt;1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

Complete chloroplast genome of Senna spectabilis (DC.) H.S. Irwin & Barneby (Fabaceae) and phylogenetic analysis

Senna spectabilis (DC.) H.S. Irwin & Barneby is a popular ornamental tree as well as a traditional medical plant in Cameroon. In this study, we sequenced and annotated the complete chloroplast genome of S. spectabilis and reconstructed the phylogenetic relationship of the tribe Cassieae. The length of the chloroplast genome was determined to be 162,754 bp, containing a pair of inverted repeats of 25,413 bp which separated by a small single-copy (SSC) region of 20,161 bp and a large single-copy (LSC) region of 91,767 bp. The cp genome encodes 128 genes, including 83 protein-coding genes, 37 tRNA genes, and eight rRNA genes. The percentage of total GC content of this genome was 35.7%. The phylogenetic analysis indicates that S. spectabilis with the sampled Senna species formed a well-supported monophyletic clade

Analysis and Optimization of Shale Oil Refinery Wastewater Treatment Process

Shale oil refinery wastewater was a kind of industrial wastewater with many types of organic matter and complex water quality. It contains a large amount of pollutants such as phenols, ammonia and oil, which is extremely harmful to the environment. Due to the increasingly stringent national discharge standards, it was difficult to treat wastewater up to standard with only one method, and multiple methods are needed to jointly treat wastewater. In this paper, the process of shale oil refinery wastewater treatment system was modified, and the “SDN+Advanced Oxidation+MBR” process was proposed to treat wastewater

Development of Isothermal Recombinase Polymerase Amplification Assay for Rapid Detection of Porcine Circovirus Type 2

Porcine circovirus virus type II (PCV2) is the etiology of postweaning multisystemic wasting syndrome (PMWS), porcine dermatitis, nephropathy syndrome (PDNS), and necrotizing pneumonia. Rapid diagnosis tool for detection of PCV2 plays an important role in the disease control and eradication program. Recombinase polymerase amplification (RPA) assays using a real-time fluorescent detection (PCV2 real-time RPA assay) and RPA combined with lateral flow dipstick (PCV2 RPA LFD assay) were developed targeting the PCV2 ORF2 gene. The results showed that the sensitivity of the PCV2 real-time RPA assay was 102 copies per reaction within 20 min at 37°C and the PCV2 RPA LFD assay had a detection limit of 102 copies per reaction in less than 20 min at 37°C. Both assays were highly specific for PCV2, with no cross-reactions with porcine circovirus virus type 1, foot-and-mouth disease virus, pseudorabies virus, porcine parvovirus, porcine reproductive and respiratory syndrome virus, and classical swine fever virus. Therefore, the RPA assays provide a novel alternative for simple, sensitive, and specific identification of PCV2

Optimized Synchronous SPWM Modulation Strategy for Traction Inverters Based on Non-Equally Spaced Carriers

The switching frequency of high-power inverters, such as those used in rail transit traction systems, is low, due to switching loss and heat dissipation limitations. This can result in considerable output voltage harmonics. This paper proposes an optimal SPWM strategy for traction inverters based on non-equally spaced carriers to address this issue. By dividing the fundamental wave cycle into regions according to the principle of three-phase, half-wave, and quarter-wave symmetry, the carrier width is symmetrically changed in each region, and a switching angle sequence is generated by comparing the fundamental wave with the non-isometric carrier. The proposed optimal modulation strategy has lower harmonic content than traditional strategies within a specific modulation range. Enhancing the inverter output waveform leads to increased torque accuracy, lowering additional losses in the motor and avoiding overheating. This results in improved performance, enhanced efficiency, and extended service life of the motor system. To further reduce voltage harmonics across the full-speed range, a multi-mode segmented synchronous modulation strategy is designed based on the optimal modulation strategy for different modulation ranges. Appropriate switching points are selected to improve the stability of the traction drive system across the full-speed range. The effectiveness of the proposed method is verified through simulation and experimental results