PKCδ is required for porcine reproductive and respiratory syndrome virus replication

AbstractProtein kinase C (PKC) that transduces signals to modulate a wide range of cellular functions has been shown to regulate a number of viral infections. Herein, we showed that inhibition of PKC with the PKC inhibitor GF109203X significantly impaired porcine reproductive and respiratory syndrome virus (PRRSV) replication. Inhibition of PKC led to virus yield reduction, which was associated with decreased viral RNA synthesis and lowered virus protein expression. And this inhibitory effect by PKC inhibitor was shown to occur at the early stage of PRRSV infection. Subsequently, we found that PRRSV infection activated PKCδ in PAMs and knockdown of PKCδ by small interfering RNA (siRNA) suppressed PRRSV replication, suggesting that novel PKCδ may play an important factor in PRRSV replication. Taken together, these data imply that PKC is involved in PRRSV infection and beneficial to PRRSV replication, extending our understanding of PRRSV replication

Comparison of Searching Behaviour of Three Evolutionary Algorithms Applied to Water Distribution System Design Optimization

Over the past few decades, various evolutionary algorithms (EAs) have been applied to the optimization design of water distribution systems (WDSs). An important research area is to compare the performance of these EAs, thereby offering guidance for the selection of the appropriate EAs for practical implementations. Such comparisons are mainly based on the final solution statistics and, hence, are unable to provide knowledge on how different EAs reach the final optimal solutions and why different EAs performed differently in identifying optimal solutions. To this end, this paper aims to compare the real-time searching behaviour of three widely used EAs, which are genetic algorithms (GAs), the differential evolution (DE) algorithm and the ant colony optimization (ACO). These three EAs are applied to five WDS benchmarking case studies with different scales and complexities, and a set of five metrics are used to measure their run-time searching quality and convergence properties. Results show that the run-time metrics can effectively reveal the underlying searching mechanisms associated with each EA, which significantly goes beyond the knowledge from the traditional end-of-run solution statistics. It is observed that the DE is able to identify better solutions if moderate and large computational budgets are allowed due to its great ability in maintaining the balance between the exploration and exploitation. However, if the computational resources are rather limited or the decision has to be made in a very short time (e.g., real-time WDS operation), the GA can be a good choice as it can always identify better solutions than the DE and ACO at the early searching stages. Based on the results, the ACO performs the worst for the five case study considered. The outcome of this study is the offer of guidance for the algorithm selection based on the available computation resources, as well as knowledge into the EA’s underlying searching behaviours

Increased Cerebral Level of P2X7R in a Tauopathy Mouse Model by PET Using [18F]GSK1482160.

Neuroinflammation plays an important role in Alzheimer's disease and primary tauopathies. The aim of the current study was to map [18F]GSK1482160 for imaging of purinergic P2X7R in Alzheimer's disease and primary tauopathy mouse models. Small animal PET was performed using [18F]GSK1482160 in widely used mouse models of Alzheimer's disease (APP/PS1, 5×FAD, and 3×Tg), 4-repeat tauopathy (rTg4510) mice, and age-matched wild-type mice. Increased uptake of [18F]GSK1482160 was observed in the brains of 7-month-old rTg4510 mice compared to wild-type mice and compared to 3-month-old rTg4510 mice. A positive correlation between hippocampal tau [18F]APN-1607 and [18F]GSK1482160 uptake was found in rTg4510 mice. No significant differences in the uptake of [18F]GSK1482160 was observed for APP/PS1 mice, 5×FAD mice, or 3×Tg mice. Immunofluorescence staining further indicated the distribution of P2X7Rs in the brains of 7-month-old rTg4510 mice with accumulation of tau inclusion. These findings provide in vivo imaging evidence for an increased level of P2X7R in the brains of tauopathy mice

Release of Matrix Metalloproteinases-2 and 9 by S-Nitrosylated Caveolin-1 Contributes to Degradation of Extracellular Matrix in tPA-Treated Hypoxic Endothelial Cells

<div><p>Intracranial hemorrhage remains the most feared complication in tissue plasminogen activator (tPA) thrombolysis for ischemic stroke. However, the underlying molecular mechanisms are still poorly elucidated. In this study, we reported an important role of caveolin-1 (Cav-1) s-nitrosylation in matrix metalloproteinase (MMP)-2 and 9 secretion from tPA-treated ischemic endothelial cells. Brain vascular endothelial cells (bEND3) were exposed to oxygen-glucose deprivation (OGD) for 2 h before adding recombinant human tPA for 6 h. This treatment induced a significant increase of MMP2 and 9 in the media of bEND3 cells and a simultaneous degradation of fibronectin and laminin β-1, the two main components of extracellular matrix (ECM). Inhibition of MMP2 and 9 with SB-3CT completely blocked the degradation of fibronectin and laminin β-1. ODG+tPA treatment led to Cav-1 shedding from bEND3 cells into the media. Notably, OGD triggered nitric oxide (NO) production and S-nitrosylationof Cav-1 (SNCav-1). Meanwhile tPA induced activation of ERK signal pathway and stimulates the secretion of SNCav-1. Pretreatment of bEND3 cells with C-PTIO (a NO scavenger) or U0126 (a specific ERK inhibitor) significantly reduced OGD-induced S-nitrosylation of Cav-1 in cells and blocked the secretion of Cav-1 and MMP2 and 9 into the media as well as the degradation of fibronectin and laminin β-1 in OGD and tPA-treated cells. These data indicate that OGD-triggered Cav-1 S-nitrosylation interacts with tPA-induced ERK activation to augment MMP2 and 9 secretion and subsequent ECM degradation, which may account for the exacerbation of ischemic blood brain barrier damage following tPA thrombolysis for ischemic stroke.</p></div

Effect of Shikonin on Spinal Cord Injury in Rats Via Regulation of HMGB1/TLR4/NF-kB Signaling Pathway

Background/Aims: Shikonin, a compound extracted from Zicao, has been demonstrated to hold anti-bacterial, anti-inflammatory, and anti-tumor activities in various diseases and it has been shown to protect human organs from injuries. However, the effect of shikonin on the recovery of spinal cord injury (SCI) remains unknown. This study was designed to estimate the potential therapeutic effect and underlying mechanism of shikonin on SCI in vivo. Methods: In the study, we used HE staining, ELISA assay, transfection assay, TUNEL assay, real time PCR and Western blot to detect the effects of shikonin on spinal cord injury in rats. Results: we showed that shikonin could promote the recovery of motor function and tissue repair after SCI treatment in rats SCI model. Moreover, we demonstrated that shikonin inhibited the spinal cord edema in SCI model of rats. According to further investigation, shikonin induced the reduction of inflammatory response through decreasing the expression levels of HMGB1, TLR4 and NF-κB after SCI injury. In addition, we also found that shikonin could suppress the apoptosis and expression of caspase-3 protein in SCI model of rats. Conclusion: Our results demonstrated that shikonin induced the recovery of tissue repair and motor function via inactivation of HMGB1/TLR4/NF-κB signaling pathway in SCI model of rats. Meanwhile, shikonin regulated the inflammation response in SCI by suppressing the HMGB1/TLR4/NF-κB signaling pathway. The described mechanism sheds novel light on molecular signaling pathway in spinal cord injury and secondary injury including inflammatory response

hBcl2 overexpression in BMSCs enhances resistance to myelin debris-induced apoptosis and facilitates neuroprotection after spinal cord injury in rats

Abstract After spinal cord injury (SCI), the accumulation of myelin debris at the lesion exacerbates cell death and hinders axonal regeneration. Transplanted bone marrow mesenchymal stem cells (BMSCs) have been proven to be beneficial for SCI repair, but they are susceptible to apoptosis. It remains unclear whether this apoptotic process is influenced by myelin debris. Here, we constructed rat BMSCs overexpressing human B-cell lymphoma 2 (hBcl2) alone (hBcl2 group), BMSCs overexpressing hBcl2 with an endoplasmic reticulum-anchored segment (hBcl2-cb) (cb group), and a negative control group (NC group) for transplantation in this study. Immunocytochemistry staining validated the successful expression of hBcl2 in BMSCs within the hBcl2 group and cb group. All BMSCs from each group exhibited the ability to phagocytize myelin debris. Nevertheless, only BMSCs derived from the hBcl2 group exhibited heightened resistance to apoptosis and maintained prolonged viability for up to 5 days when exposed to myelin debris. Notably, overexpression of hBcl2 protein, rather than its endoplasmic reticulum-anchored counterpart, significantly enhanced the resistance of BMSCs against myelin debris-induced apoptosis. This process appeared to be associated with the efficient degradation of myelin debris through the Lamp1+ lysosomal pathway in the hBcl2 group. In vivo, the hBcl2 group exhibited significantly higher numbers of surviving cells and fewer apoptotic BMSCs compared to the cb and NC groups following transplantation. Furthermore, the hBcl2 group displayed reduced GFAP+ glial scarring and greater preservation of NF200+ axons in the lesions of SCI rats. Our results suggest that myelin debris triggers apoptosis in transplanted BMSCs, potentially elucidating the low survival rate of these cells after SCI. Consequently, the survival rate of transplanted BMSCs is improved by hBcl2 overexpression, leading to enhanced preservation of axons within the injured spinal cord

Kirigami-inspired multiscale patterning of metallic structures via predefined nanotrench templates.

Reliable fabrication of multiscale metallic patterns with precise geometry and size at both the nanoscale and macroscale is of importance for various applications in electronic and optical devices. The existing fabrication processes, which usually involve film deposition in combination with electron-beam patterning, are either time-consuming or offer limited precision. Inspired by the kirigami, an ancient handicraft art of paper cutting, this work demonstrates an electron-beam patterning process for multiscale metallic structures with significantly enhanced efficiency and precision. Similar to the kirigami, in which the final pattern is defined by cutting its contour in a paper and then removing the unwanted parts, we define the target multiscale structures by first creating nanotrench contours in a metallic film via an electron-beam-based process and then selectively peeling the separated film outside the contours. Compared with the conventional approach, which requires the exposure of the whole pattern, much less exposure area is needed for nanotrench contours, thus enabling reduced exposure time and enhanced geometric precision due to the mitigated proximity effect. A theoretical model based on interface mechanics allows a clear understanding of the nanotrench-assisted selective debonding behaviour in the peeling process. By using this fabrication process, multiscale metallic structures with sub-10-nm up to submillimetre features can be reliably achieved, having potential applications for anti-counterfeiting and gap-plasmon-enhanced spectroscopy

tPA promotes MMP2 and 9-mediated fibronectin and laminin β-1 degradation in OGD-treated endothelial cells.

<p><b><i>A)</i></b> The levels of fibronectin (Fb) and laminin β-1 (LMN β-1) significantly decreased in bEND3 that were first exposed to 2-h OGD and followed by tPA treatment for 6 h (O+t) compared with untreated cells (Ctrl) or the cells that were treated by OGD or tPA alone. Upper panel: representative immunoblots, β-actin served as loading control; Bottom panel: band intensity was quantitated after normalization to β-actin and the data were expressed as mean ± SD, *<i>p</i> < 0.05, ANOVA, N = 4. <b><i>B)</i></b> Gelatin zymography analysis showed that OGD+tPA treatment (O+t) significantly increased MMP2 and 9 levels in the conditioned media. Upper panel: representative gelatin zymograms; Bottom panel: band intensity was quantitated and the data were expressed as mean ± SD, *<i>p</i> < 0.05, ANOVA, N = 4. Std: human MMP2 and 9 standards. <b><i>C)</i></b> SB-3CT completely blocked OGD+tPA (O+t)-induced degradation of fibronectin (Fb) and laminin β-1 (LMN β-1). Left panel: representative immunoblots; Right panel: band intensity was quantitated after normalization to β-actin and the data were expressed as mean ± SD, ANOVA, N = 4.</p