4-1BB(CD137) is a member of the tumor necrosis factor receptor superfamily and provides a costimulatory signal by interaction with 4-1BB ligand expressed on antigen-presenting cells. The expression of 4-1BB is known to be activationdependent. Here, we investigated the transcriptional machinery required for T cell receptor (TCR) activation-dependent induction of 4-1BB expression in CD3-CEM cells treated with phorbol myristate acetate and ionomycin. Using 5P P-deletion constructs of 4-1BB promoter in luciferase reporter assays, we demonstrated that the transcriptional elements mediating 4-1BB upregulation were located in the region between V V0.9 and V V1.1 kb from the translational start site. Characterization of these sites by electrophoretic mobility shift assay and sitedirected mutagenesis revealed that nuclear factor U UB (NF-U UB) and activating protein-1 (AP-1) are involved. MEK and c-Jun N-terminal kinase-1 activity was required for activation-dependent 4-1BB upregulation. Thus, NF-U UB and AP-1 are involved in the TCR stimulation-dependent transcriptional regulation of the 4-1BB promoter. ß

Two-dimensional quantitative nuclear magnetic resonance (2D qNMR)-based metabolomics was performed to understand characteristic metabolic profiles in different aging regimes (crust from dry-aged beef, inner edible flesh of dry-aged beef, and wet-aged beef striploin) over 4 weeks. Samples were extracted using 0.6 M perchlorate to acquire polar metabolites. Partial least squares-discriminant analysis showed a good cumulative explained variation (R2 = 0.967) and predictive ability (Q2 = 0.935). Metabolites of crust and aged beef (dry- and wet-aged beef) were separated in the first week and showed a completely different aspect in the second week via NMR-based multivariable analyses. Moreover, NMR-based multivariable analyses could be used to distinguish the method, degree, and doneness of beef aging. Among them, the crust showed more unique metabolic changes that accelerated proteolysis (total free amino acids and biogenic amines) and inosine 5′-monophosphate depletion than dry-aged beef and generated specific microbial catabolites (3-indoxyl sulfate) and γ-aminobutyric acid (GABA), while asparagine, glutamine, tryptophan, and glucose in the crust were maintained or decreased. Compared to the crust, dry-aged beef showed similar patterns of biogenic amines, as well as bioactive compounds and GABA, without a decrease in free amino acids and glucose. Based on these results, the crust allows the inner dry-aged beef to be aged similarly to wet-aged beef without microbial effects. Thus, 2D qNMR-based metabolomic techniques could provide complementary information about biochemical factors for beef aging.

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The objective of this study was to investigate the effects of canola and flaxseed oils on the physicochemical properties and sensory quality of emulsion-type sausage made from spent layer meat. Three types of sausage were manufactured with different fat sources: 20% pork back fat (CON), 20% canola oil (CA) and 20% flaxseed oil (FL). The pH value of the CA was significantly higher than the others (p<0.05). The highest water holding capacity was also presented for CA; in other words, CA demonstrated a significantly lower water loss value among the treatments (p<0.05). CA had the highest lightness value (p<0.05). However, FL showed the highest yellowness value (p<0.05) because of its own high-density yellow color. The texture profile of the treatments manufactured with vegetable oils showed higher values than for the CON (p<0.05); furthermore, CA had the highest texture profile values (p<0.05) among the treatments. The replacement of pork back fat with canola and flaxseed oils in sausages significantly increased the omega-3 fatty acid content (p<0.05) over 15 to 86 times, respectively. All emulsion sausages containing vegetable oil exhibited significantly lower values for saturated fatty acid content and the omega-6 to omega-3 ratios compared to CON (p<0.05). The results show that using canola or flaxseed oils as a pork fat replacer has a high potential to produce healthier products, and notably, the use of canola oil produced characteristics of great emulsion stability and sensory quality.