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Abstract: Nitric oxide (NO) causes S-glutathiolation of the reactive cysteine-674 in the sarcoplasmic/endoplasmic reticulum Ca2+ ATPase (SERCA), thus increasing SERCA activity, and inhibiting Ca2+ influx and migration of vascular smooth muscle cells (VSMC). Because increased VSMC migration contributes to accelerated neointimal growth and atherosclerosis in diabetes, the effect of culture of VSMC in high glucose (HG) was determined. Rat aortic VSMC were exposed to normal (5.5 mmol/L) or high (25 mmol/L) glucose for 3 days, and serum-induced cell migration during 6 h into a wounded cell monolayer was measured 5 min after adding the NO donor S-nitroso-N-acetylpenicillamine (SNAP) or 24 h after interleukin-1β (IL-1β) to express inducible nitric oxide synthase (iNOS). In normal glucose, SNAP or IL-1β significantly inhibited migration in cells infected with adenovirus to express GFP or SERCA wild type (WT), but not with a C674S SERCA mutant. After HG, NO failed to inhibit migration, nor did it decrease calcium-dependent association of calmodulin with calcineurin, indicating that NO failed to decrease intracellular calcium levels via SERCA. In contrast, overexpression of SERCA WT, but not the SERCA C674S mutant, preserved the ability for NO to inhibit migration despite exposing the cells to HG. The antioxidant, Tempol, or overexpression of superoxide dismutase also prevented the effects of HG. Further studies showed that both biotinylated-iodoacetamide and NO-induced biotinylated glutathione labeling of SERCA C674 were decreased by HG, and a sequence-specific sulfonic acid antibody detected oxidation of the C674 SERCA thiol. These results indicate that failure of NO to inhibit migration in VSMC exposed to HG is due to oxidation of the SERCA reactive cysteine-674. [Copyright &y& Elsevier]

Receptor interacting protein 1 (RIP1) plays important roles not only in cell-death pathways but also in host innate immune responses. In the present study, a RIP1 ortholog named Lc-RIP1 was cloned and characterized in large yellow croaker (Larimichthys crocea). The open reading frame (ORF) of Lc-RIP1 is 2,046 bp, encoding a protein of 681 amino acids (aa), with an N-terminal kinase domain, an RHIM domain, and a C-terminal death domain. Subcellular localization analysis revealed that Lc -RIP1 was a cytosolic protein, which was broadly expressed in examined tissues/organs, and could be up-regulated under poly I:C, LPS, PGN, and Pseudomonas plecoglossicida stimulation in vivo based on qRT-PCR analysis. Notably, Lc -RIP1 could induce NF-κB, but not IRF3, IRF7 or type I IFN promoter activation. In addition, Lc -RIP1 overexpression could enhance Lc -MAVS, Lc -TRAF3, and Lc -TRAF6 mediated NF-κB promoter activation, and also Lc -TRIF and Lc -MAVS mediated IRF3 promoter activation, whereas suppress Lc -TRIF mediated NF-κB and type I IFN promoter activation, as well as Lc -TRAF3 and Lc -IRF3 mediated IRF3 promoter activation, Lc -IRF3 mediated type I IFN promoter activation and Lc -IRF7 mediated IRF7 promoter activation. These results collectively indicated that Lc -RIP1 function importantly in regulation of host innate immune signaling. • Lc-RIP1 can be induced by both viral and bacterial PAMPs stimulation. • Lc -RIP1 induces NF-κB, but not IRF3, IRF7 or type I IFN promoter activation. • Lc -RIP1 enhances MAVS, TRAF3, and TRAF6 mediated NF-κB activation, and also TRIF and MAVS mediated IRF3 activation. • Lc -RIP1 suppresses TRIF mediated NF-κB activation, and also TRAF3 and IRF3 mediated IRF3 activation. [ABSTRACT FROM AUTHOR]

At present, stem cell transplantation has a significant therapeutic effect on spinal cord injury (SCI), however, it is still challenging for the seed cells selection. In this study, in order to explore cells with wide neural repair potentials, we selected the pluripotent stem cells multilineage-differentiating stress-enduring (Muse) cells, inducing the in vitro differentiation of human Muse cells into neural precursor cells (Muse-NPCs) by applying neural induction medium. Here, we found induced Muse-NPCs expressed neural stem cell markers Nestin and NCAM, capable of differentiating into three types of neural cells (neuron, astrocyte and oligodendrocyte), and have certain biological functions. When Muse-NPCs were transplanted into rats suffering from T10 SCI, motor function was improved. These results provide an insight for application of Muse-NPCs in cell therapy or tissue engineering for the repair of SCI in future. • Muse cells can be induced into neural precursor cells in vitro. • Muse-NPCs different into neurons, astrocytes and oligodendrocytes with their own functions in vitro. • Muse-NPCs promote the restoration of motor function in SCI rats. [ABSTRACT FROM AUTHOR]

As an adaptor protein functions essentially in the activation of NF-κΒ and MAPK signaling pathways mediated by NOD1 and NOD2, RIP2 plays important roles in the host innate immune responses. In the present study, the RIP2 ortholog termed Lc- RIP2 was identified and characterized in large yellow croaker (Larimichthys crocea). It was revealed that Lc- RIP2 is consisted of an open reading frame (ORF) of 1695 bp, encoding a protein of 564 aa, with an N-terminal kinase domain and a C-terminal caspase activation and recruitment domain (CARD). Subcellular localization assays demonstrated that Lc- RIP2 was a cytosolic protein, which was broadly distributed in the examined tissues/organs, and could be induced in response to poly I:C, LPS, PGN, and Pseudomonas plecoglossicida stimulations in vivo according to qRT-PCR analysis. Notably, Lc -RIP2 overexpression in vitro was sufficient to abolish SVCV proliferation in EPC cells, and could significantly induce the activation of NF-κB, IRF3, IRF7, and IFN1 promoters. In addition, luciferase assays found that Lc- RIP2 could cooperate with Lc -MAVS, Lc -TRAF3, Lc -TRAF6, Lc -IRF3, and Lc -IRF7 in NF-κB activation, associate with Lc -TRIF, Lc -MAVS, Lc -TRAF3, Lc -IRF3, and Lc -IRF7 in IRF3 activation, enhance Lc -TRIF, Lc -MAVS, Lc -TRAF3, and Lc -TRAF6 mediated IRF7 activation, and Lc -IRF3 mediated IFN1 activation, whereas suppress NF-κB activation when co-expressed with Lc -TRIF. Co-immunoprecipitation (Co-IP) assays also demonstrated that Lc -RIP2 interacts separately with Lc -TRIF, Lc -MAVS, Lc -TRAF3, Lc -TRAF6, Lc -IRF3, and Lc -IRF7. It is thus collectively indicated that Lc -RIP2 function dominantly in the regulation of the host innate immune signaling. • Lc -RIP2 induces NF-κB, IRF3, IRF7, and type I IFN promoter activation. • Lc -RIP2 overexpression suppresses virus proliferation. • Lc -RIP2 interacts with TRIF, MAVS, TRAF3, TRAF6, IRF3, and IRF7. • Lc -RIP2 associates with IRF3 in IRF3 and type I IFN promoter activation. • Lc -RIP2 enhances MAVS, TRAF3, TRAF6, IRF3, and IRF7 mediated NF-κB activation, whereas suppresses TRIF mediated NF-κB activation. [ABSTRACT FROM AUTHOR]

Highlights: [•] A novel GATA4 mutation, p.C271S, was identified in a large family with DCM. [•] The mutation co-segregated with DCM in the family and was absent in 200 controls. [•] The altered amino acid was completely conserved evolutionarily among species. [•] The mutant was associated with significantly decreased transcriptional activity. [•] The mutant remarkably reduced the synergistic activation between GATA4 and NKX2-5. [ABSTRACT FROM AUTHOR]