Targeted inCITE-Seq Analysis Identifies the Loss of Nuclear TDP-43 in Endothelium as a Mediator of Blood Brain Barrier Signaling Pathway Dysfunction in Neurodegeneration.

Despite the importance of the endothelium in the regulation of the blood brain barrier (BBB) in aging and neurodegenerative disease, difficulties in extracting endothelial cell (EC) nuclei have limited analysis of these cells. In addition, nearly all Amyotrophic Lateral Sclerosis (ALS) and Frontotemporal Degeneration (FTD), and a large portion of Alzheimer's Disease (AD) exhibit neuronal TDP-43 aggregation, leading to loss of nuclear function, but whether TDP-43 is similarly altered in human BBB ECs is unknown. Here we utilize a novel technique for the enrichment of endothelial and microglial nuclei from human cortical brain tissues, combined with inCITE-seq, to analyze nuclear proteins and RNA transcripts in a large cohort of healthy and diseased donors. Our findings reveal a unique transcriptional signature in nearly half of the capillary endothelial cells across neurodegenerative states, characterized by reduced levels of nuclear ß-Catenin and canonical downstream genes, and an increase in TNF/NF-kB target genes. We demonstrate that this does not correlate with increased nuclear p65/NF-kB, but rather a specific loss of nuclear TDP-43 in these disease associated ECs. Comparative analysis in animal models with targeted disruption of TDP-43 shows that this is sufficient to drive these transcriptional alterations. This work reveals that TDP-43 is a critical governor of the transcriptional output from nuclear p65/NF-kB, which has paradoxical roles in barrier maintenance and also barrier compromising inflammatory responses, and suggests that disease specific loss in ECs contributes to BBB defects observed in the progression of AD, ALS and FTD.

Splice factor polypyrimidine tract-binding protein 1 (Ptbp1) primes endothelial inflammation in atherogenic disturbed flow conditions.

NF-κB-mediated endothelial activation drives leukocyte recruitment and atherosclerosis, in part through adhesion molecules Icam1 and Vcam1. The endothelium is primed for cytokine activation of NF-κB by exposure to low and disturbed blood flow (LDF)but the molecular underpinnings are not fully understood. In an experimental in vivo model of LDF, platelets were required for the increased expression of several RNA-binding splice factors, including polypyrimidine tract binding protein (Ptbp1). This was coordinated with changes in RNA splicing in the NF-κB pathway in primed cells, leading us to examine splice factors as mediators of priming. Using Icam1 and Vcam1 induction by tumor necrosis factor (TNF)-α stimulation as a readout, we performed a CRISPR Cas9 knockout screen and identified a requirement for Ptbp1 in priming. Deletion of Ptbp1 had no effect on cell growth or response to apoptotic stimuli, but reversed LDF splicing patterns and inhibited NF-κB nuclear translocation and transcriptional activation of downstream targets, including Icam1 and Vcam1. In human coronary arteries, elevated PTBP1 correlates with expression of TNF pathway genes and plaque. In vivo, endothelial-specific deletion of Ptbp1 reduced Icam1 expression and myeloid cell infiltration at regions of LDF in atherosclerotic mice, limiting atherosclerosis. This may be mediated, in part, by allowing inclusion of a conserved alternative exon in Ripk1 leading to a reduction in Ripk1 protein. Our data show that Ptbp1, which is induced in a subset of the endothelium by platelet recruitment at regions of LDF, is required for priming of the endothelium for subsequent NF-κB activation, myeloid cell recruitment and atherosclerosis.

Review on grain size effects on thermal conductivity in ZnO thermoelectric materials.

Thermoelectric materials have recently attracted a lot of attention due to their ability to convert waste heat into electricity. Based on the extensive research in this area, the nanostructuring approach has been viewed as an effective strategy for increasing thermoelectric performance. This approach focuses on the formation and growth of the superfine, pure and uniform grain size. Since the grain size has a strong influence on the thermal conductivity, this can be reduced by increasing the phonon scattering at grain boundaries and refining the grain sizes. Therefore, this review aims to discuss the mechanism of reduction in thermal conductivity in small-grain zinc oxide (ZnO) and the optimization techniques for obtaining ZnO nanoparticles with desirably low thermal conductivity and excellent thermoelectric performance.

Dual-wavelength, mode-locked erbium-doped fiber laser employing a graphene/polymethyl-methacrylate saturable absorber.

Mode-locked fiber laser incorporating a saturable absorber is an attractive configuration due to its stability and simple structure. In this work, we demonstrate a dual-wavelength passively mode-locked erbium-doped fiber laser employing a graphene/polymethyl-methacrylate saturable absorber. A laser resonator is developed based on dual cavity architecture with unidirectional signal oscillation, which is connected by a fiber branch sharing a common gain medium and saturable absorber. Dual wavelength mode-locked fiber lasers are observed at approximately 1530 and 1560 nm with 22.6 mW pump power threshold. Soliton pulse circulates in the laser cavity with pulse duration of 900 and 940 fs at shorter and longer wavelengths, respectively. This work presents a viable option in developing a low threshold mode-locked laser source with closely spaced dual wavelength femtosecond pulses in the C-band wavelength region.