Multi-functional regulation of cGAS by the nuclear localization signal2 (NLS2) motif: Nuclear localization, enzyme activity and protein degradation.

Cyclic GMP-AMP synthase (cGAS), which recognizes double-stranded DNA (dsDNA) and activates the innate immune system, is mainly localized in the cytosol, but also shows nuclear localization. Here, we sought to determine the role of nuclear cGAS by mutating known nuclear localization signal (NLS) motifs in cGAS and assessing its functionality by monitoring phosphorylation of the downstream target, interferon regulatory factor-3 (IRF3). Interestingly, NLS2-mutated cGAS failed to promote phosphorylation of IRF3, reflecting the loss of its ability to produce cyclic GMP-AMP (cGAMP). We further found that insertion of an NLS from SV40 large T antigen could not restore this loss of activity, indicating that this loss was attributable to the mutation of NLS2 itself, but not dependent on the inability of cGAS to enter the nucleus. NLS2-mutant cGAS protein also showed decreased stability dependent on polyubiquitination, an effect that was independent of both its loss of catalytic function and its inability to enter into the nucleus. Collectively, these findings indicate that the NLS2 motif of cGAS is not only involved in regulating the subcellular localization of cGAS protein but also influences its stability and enzymatic activity through independent mechanisms, highlighting the novel roles of NLS2 in regulating the intracellular functions of cGAS.

A Systematic Review on Recent Trends, Challenges, Privacy and Security Issues of Underwater Internet of Things.

Owing to the hasty growth of communication technologies in the Underwater Internet of Things (UIoT), many researchers and industries focus on enhancing the existing technologies of UIoT systems for developing numerous applications such as oceanography, diver networks monitoring, deep-sea exploration and early warning systems. In a constrained UIoT environment, communication media such as acoustic, infrared (IR), visible light, radiofrequency (RF) and magnet induction (MI) are generally used to transmit information via digitally linked underwater devices. However, each medium has its technical limitations: for example, the acoustic medium has challenges such as narrow-channel bandwidth, low data rate, high cost, etc., and optical medium has challenges such as high absorption, scattering, long-distance data transmission, etc. Moreover, the malicious node can steal the underwater data by employing blackhole attacks, routing attacks, Sybil attacks, etc. Furthermore, due to heavyweight, the existing privacy and security mechanism of the terrestrial internet of things (IoT) cannot be applied directly to UIoT environment. Hence, this paper aims to provide a systematic review of recent trends, applications, communication technologies, challenges, security threats and privacy issues of UIoT system. Additionally, this paper highlights the methods of preventing the technical challenges and security attacks of the UIoT environment. Finally, this systematic review contributes much to the profit of researchers to analyze and improve the performance of services in UIoT applications.

Comparison of mid- to long-term clinical outcomes between anatomical testing and usual care in patients with suspected coronary artery disease: A meta-analysis of randomized trials.

BACKGROUND: Controversies remain regarding clinical outcomes following initial strategies of coronary computed tomography angiography (CCTA) vs usual care with functional testing in patients with suspected coronary artery disease (CAD). HYPOTHESIS: CCTA as initial diagnostic strategy results in better mid- to long-term outcomes than usual care in patients with suspected CAD. METHODS: We searched PubMed, Embase, and Cochrane Library for randomized controlled trials comparing clinical outcomes during ≥6 months' follow-up between initial anatomical testing by CCTA vs usual care with functional testing in patients with suspected CAD. Occurrence of all-cause mortality, nonfatal myocardial infarction (MI), and major adverse cardiovascular events (MACE), and use of invasive coronary angiography and coronary revascularization, were compared between the 2 diagnostic strategies. RESULTS: Twelve trials were included (20 014 patients; mean follow-up, 20.5 months). Patients undergoing CCTA as initial noninvasive testing had lower risk of nonfatal MI compared with those treated with usual care (risk ratio [RR]: 0.70, 95% confidence interval [CI]: 0.52-0.94, P = 0.02). There was a tendency for reduced MACE following initial CCTA strategy, but not for risk of all-cause mortality. Compared with functional testing, the CCTA strategy increased use of invasive coronary angiography (RR: 1.53, 95% CI: 1.12-2.09, P = 0.007) and coronary revascularization (RR: 1.49, 95% CI: 1.11-2.00, P = 0.007). CONCLUSIONS: Anatomical testing with CCTA as the initial noninvasive diagnostic modality in patients with suspected CAD resulted in lower risk of nonfatal MI than usual care with functional testing, at the expense of more frequent use of invasive procedures.

Dual oxidase 2 is essential for house dust mite-induced pro-inflammatory cytokine production in human keratinocytes.

House dust mites (HDMs) are known to trigger chronic inflammation through Toll-like receptors (TLRs) and their signalling cascades. In this study, we found that TLR2 ligation by HDMs induced the activation of dual oxidase 2 (Duox2) and nuclear factor-κB (NF-κB), leading to the production of pro-inflammatory cytokines in human keratinocytes. Stimulation of human keratinocytes with HDMs resulted in increases in interleukin-8 (IL-8) and chemokine (C-C motif) ligand 20 (CCL20) levels. However, pro-inflammatory cytokine production was abolished in keratinocytes transfected with TLR2 siRNA, indicating that HDM-induced cytokine production was mediated via TLR2 signalling. We also examined the function of Duox1/2 isozymes, which are primarily expressed in keratinocytes, in HDM-mediated pro-inflammatory cytokine production. Human keratinocytes transfected with control siRNA or Duox1 siRNA showed no inhibition of IL-8 or CCL20 production in response to HDMs, whereas the silencing of Duox2 expression resulted in a failure to induce cytokine production. Moreover, the phosphorylation and nuclear localization of RelA/p65, a component of NF-κB, were induced by HDMs in human keratinocytes. Transfection of human keratinocytes with TLR2 siRNA or Duox2 siRNA resulted in the complete abolishment of RelA/p65 nuclear localization in response to HDMs. Taken together, these results indicate that the HDM-dependent TLR2-Duox2 signalling axis indeed promotes NF-κB activation, which induces IL-8 and CCL20 production and mediates epidermal keratinocyte inflammation.

Testosterone stimulates Duox1 activity through GPRC6A in skin keratinocytes.

Testosterone is an endocrine hormone with functions in reproductive organs, anabolic events, and skin homeostasis. We report here that GPRC6A serves as a sensor and mediator of the rapid action of testosterone in epidermal keratinocytes. The silencing of GPRC6A inhibited testosterone-induced intracellular calcium ([Ca(2+)]i) mobilization and H2O2 generation. These results indicated that a testosterone-GPRC6A complex is required for activation of Gq protein, IP3 generation, and [Ca(2+)]i mobilization, leading to Duox1 activation. H2O2 generation by testosterone stimulated the apoptosis of keratinocytes through the activation of caspase-3. The application of testosterone into three-dimensional skin equivalents increased the apoptosis of keratinocytes between the granular and stratified corneum layers. These results support an understanding of the molecular mechanism of testosterone-dependent apoptosis in which testosterone stimulates H2O2 generation through the activation of Duox1.

Cell chip with a thiolated chitosan self-assembled monolayer to detect the effects of anticancer drugs on breast normal and cancer cells.

Cell-based chips are an effective in vitro analysis tool; however, the sensitivity of the cell chip to biomaterials is high, which is crucial for immobilizing cells on the electrode surface without conductivity. In this study, we report on a cell chip with a thiolated chitosan monolayer that was easy to fabricate, highly adhesive to cells, and enhanced electrochemical signals. Thiolated chitosan containing thiol groups was synthesized and self-assembled on a gold electrode to immobilize cells, and showed superior electrochemical performance to that of poly-l-lysine and collagen. Cyclic voltammetry (CV) was performed to distinguish the redox characteristics of normal (HMEC) and breast cancer cells (MCF-7); then, two anticancer drugs (doxorubicin and cyclophosphamide) were added to the cell cultures to analyze their effects on the redox environment of normal and cancer cells derived from the same origin. As a result, the CV cathode peaks decreased differently with respect to the cell line (normal and cancer) and anticancer drug, which was validated by a conventional MTT viability assay. Hence, the proposed cell chip with a thiolated chitosan modified layer could be used in various fields, including discriminating normal from cancer cells, to evaluating the efficiency of newly developed drugs, and to assessing cytotoxicity of various chemicals.

Nanoscale film fabrication of various peptides on neural stem cell chip.

Modification of peptide on the electrode surface is very important issue for achieving valuable information from cell chip. In this study, various kinds of cysteine-containing peptide were fabricated on the electrode surface to enhance the electrochemical signals, cell spreading, and proliferation of rat neural stem cells. Different kinds of lysine-rich and RGD peptides were self-assembled on the gold nanoparticle modified ITO surfaces via strong Au-S chemical bond, followed by seeding neural stem cells (NE-4C) on its surface. As a result, K-MAP-C peptide consists of the quadruple branches of lysine chains and cysteine terminal showed outstanding characteristics respect to the improvement of redox signals, cell spreading and proliferation on electrode surface. Hence, our stem cell chip composed of lysine-rich peptide modified electrode can be usefully applied as efficient stem cell research tool.