KCa2.3 expression was significantly reduced in mesenteric resistance arteries in AMPKα2 knockout mice when compared with littermate control mice. Furthermore, in high-fat diet fed mice, 2-week treatment with AICAR restored endothelial KCa2.3 expression in mesenteric resistance arteries with improved endothelial dysfunction. Our results demonstrate that activation of AMPK upregulates KCa2.3 channel expression through the ERK5-MEF2-KLF2/4 signaling pathway in vascular endothelium, which contributes to benefits through KCa2.3-mediated EDH-type vasodilation in mesenteric resistance arteries.Advanced glycation end products (AGEs) are a heterogenous group of glycation adducts on amino acids produced with sugars or dicarbonyls. Intracellular inflammation triggered by binding of AGEs to receptor for AGEs (RAGE) is linked to some chronic diseases. Here, we established a competitive assay format to comprehensively quantify AGEs which bound to RAGE. RAGE-binding activities of sugar- and dicarbonyl-derived AGEs were correlated with oxidative stress in cultured cells generated by the respective AGEs, suggesting that this would be a promising method for evaluating AGEs which could affect cellular functions despite limited information on individual glycation adducts.Antifreeze proteins (AFP) play an important role in cellular survival at sub-zero temperatures. This study assessed the effect of AFP type I or III in semen extender (TRIS-egg yolk) for ram sperm cryopreservation. Pooled semen of four rams were allocated into five treatments Control (CONT, without AFP); AFP Type I [0.1 (AFPI-0.1) or 0.5 (AFPI-0.5) μg/mL]; or III [0.1 (AFPIII-0.1) or 0.5 (AFPIII-0.5) μg/mL], and then frozen in six replicates. Terephthalic in vivo Treatments affected kinetic parameters, plasma membrane integrity and morphology (P 0.05) were observed in hypoosmotic test, sperm acrosome status, mitochondrial activity, chromatin condensation, perivitelline membrane binding rate and lipoperoxidation. In conclusion, the use of AFP, predominantly type I, may increase sperm cell protection during cryopreservation, with no adverse effect on potential fertilization capacity or increase in reactive oxygen species, being a potential cryoprotectant to ram sperm.Spontaneous development of atopic dermatitis (AD) in NC/Nga (NC) mice has been attributed to a deficiency in invariant NK T (iNKT) cells. To elucidate the precise role of iNKT cells in AD development of NC mice, we employed two distinct murine models of iNKT cell over-representation Vβ8 TCR congenic and Vα14 TCR transgenic NC mice. We found that Vα14 TCR transgenic (Vα14Tg) but not Vβ8 TCR congenic (Vβ8Cg) NC mice exhibited reduced AD development, which was attributed to both quantitative and qualitative changes in iNKT cells such as a biased expansion of the double-negative iNKT subset. Adoptive transfer experiments confirmed that iNKT cells from Vα14Tg mice but not from Vβ8Cg mice were responsible for protecting NC mice from AD development. Double-negative iNKT cells from Vα14Tg NC mice showed a T helper type-1‒dominant cytokine profile, which may account for the expansion of CD4+ regulatory T cells and memory-type CD8+ T cells. Furthermore, the adoptive transfer of CD8+ T cells from Vα14Tg NC mice into AD-susceptible wild-type NC mice suppressed AD in recipient NC mice. Taken together, our results have identified double-negative iNKT cells as promising cellular targets to prevent AD pathogenesis.The mechanism of high-grade transformation in gastrointestinal stromal tumors (GISTs) remains to be clarified. We aim to discover the key progression events by studying biphasic GISTs. The study group included 101 GISTs. Nineteen of these had been screened from 263 GISTs to represent the early stage of GIST high-grade transformation, characterized by juxtaposed low-grade and high-grade regions in the same tumor (so-called biphasic GISTs). Mutational analyses, fluorescence in situ hybridization (FISH), NanoString analyses, telomere analysis, and gene expression profiling were carried out, followed by in silico analyses, cell line study, and immunohistochemical validation. Using gene expression analysis, downregulation of SFRP1 was revealed to be the main event in GIST high-grade transformation (p = 0.013), accompanied by upregulation of EZH2. In silico analyses revealed that downregulation of SFRP1 was a common feature in GIST progression across several different series. Immunohistochemically, the expression of SFRP1 was validated to be significantly lower in high-grade GISTs (WHO risk group 3a or higher) than in low-grade GISTs (p less then 0.001), and attenuation/loss of SFRP1 was associated with GIST tumor progression (p less then 0.001). By NanoString and FISH analyses, chromosomal 9/9p loss was the only recurrent large-scale chromosome aberration in biphasic GISTs, with a correlation with SFRP1 downregulation. Subclones containing chromosome 9/9p loss could be appreciated in the low-grade parts of biphasic GISTs. TP53 mutation, RB1 loss, KIT/PDGFRA mutation, and alternative lengthening of telomeres did not play a significant role in GIST high-grade transformation. In conclusion, high-grade transformation of GISTs features SFRP1 downregulation and chromosome 9/9p loss.Recombinant hybrid antibodies are commonly used in antigen-targeting assays to reduce the immunogenic potential associated with using classic mouse antibodies in other species. The DEC205 receptor has become an attractive target due to its effectiveness in activating the immune response and is considered a promising vaccination target. The aim of this study was to produce a fully chimeric mouse x pig anti-porcine DEC205 recombinant antibody (rAb). Based on a mouse anti-porcine DEC205 monoclonal antibody (mAb), we designed and expressed a chimeric mouse x pig rAb using the Expi293f system. The resulting rAb maintained the recognition capacity of the native mouse mAb toward the porcine DEC205 receptor, as evidenced by western blot analysis. By using flow cytometry, we evaluated the ability of the rAb to recognize DEC205+ dendritic cells. In conclusion, the chimeric mouse x pig anti-DEC205 rAb can be used in antigen-targeting assays as a vaccination strategy in pigs.