thrombin cleavage site  (Thermo Fisher)

Journal: Cell Reports Medicine

Article Title: Spleen-targeted neoantigen mRNA vaccine induces ISG15 + CD8 + T cell-mediated tertiary lymphoid structure formation in hepatocellular carcinoma

doi: 10.1016/j.xcrm.2026.102754

Figure Lengend Snippet: GZMA-F2R interaction mediates STNvac efficacy through T cell activation (A) Bubble plot showing ligand-receptor pairs between ISG15 + CD8 + T cells and APCs (B cells, CD4 + T cells, and DCs). (B) Violin plots showing F2R expression across T cell subsets and GZMA expression in APCs. (C and D) Therapeutic evaluation of STNvac co-administration with F2R antagonist (F2RA, SCH79797 ) ( n = 7 mice per group). (C) Bioluminescence images showing tumor burden in orthotopic HCC-bearing mice under the indicated treatments. (D) Survival curves of mice in different groups. (E and F) Multicolor immunofluorescence staining of intratumoral Ki67 + CD69 + ISG15 + CD8 + T cells. (E) Representative images. Scale bars, 20 μm. (F) Quantitative analysis of positive cell density in five randomly selected areas per tumor section. (G and H) Activation of human HCC TILs through the GZMA-F2R interaction. (G) Schematic illustration of the treatment schedule. (H) Quantitative analysis of 41BB + CD3 + CD8 + T cells after 24 h of GZMA stimulation ( n = 2 biological replicates, each analyzed in triplicate). Statistics: one-way ANOVA for (F) and (H); log rank (Mantel-Cox) test for (D). Mean ± SD. Significance levels: ∗p < 0.05, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. See also and .

Article Snippet: To evaluate the impact of the GZMA-F2R signaling on CD8 + TIL activation, the in vitro-expanded TILs were seeded in 24-well plates, pre-treated with the F2R antagonist SCH79797 (MCE, 5 μM), and subsequently stimulated with recombinant GZMA (MCE, 0.05 ng/mL).

Techniques: Activation Assay, Expressing, Multicolor Immunofluorescence Staining

Journal: Science Advances

Article Title: Hemostatic B-knob–triggered microgels (BK-TriGs) to address bleeding in neonates

doi: 10.1126/sciadv.ady7698

Figure Lengend Snippet: ( A ) Schematic representation of the experimental timeline: neonatal fibrinogen injection, followed by microparticle injection, liver laceration, and subsequent blood loss monitoring over 10 min. ( B ) Time course of blood loss (grams of blood per gram of animal weight) in mice treated with saline or varying doses of BK-TriGs (10, 15, and 20 mg/kg), demonstrating dose-dependent hemostatic effects. ( C ) Total blood loss analysis showing significant reduction with BK-TriGs at 15 mg/kg compared to saline ( P < 0.001) and highlighting increased blood loss at 20 mg/kg ( P < 0.0001). ( D ) Comparative blood loss over time for mice treated with NB-ULCs, AK-ULCs, and BK-TriGs at 15 mg/kg, showing superior performance of BK-TriGs. ( E ) Total blood loss for the different particle treatments, indicating the significant efficacy of BK-TriGs ( P < 0.05) compared to other groups. Data are presented as the means ± SD.

Article Snippet: In each 50-μl reaction, 45.25 μl of plasma was mixed with 1 μl of Alexa Fluor 488–labeled human fibrinogen (10 μg/ml final; Thermo Fisher Scientific), 1.25 μl of CaCl 2 (200 mM stock; final 5 mM), and 2.5 μl of human thrombin (10 U/ml; Enzyme Research Laboratories, US) to initiate polymerization [thrombin (final 0.5 U/ml)].

Techniques: Injection, Saline

Journal: Gels

Article Title: Origin of the High Variability in Sol–Gel Phase Transitions: The Agar Gelation Model

doi: 10.3390/gels12040304

Figure Lengend Snippet: ( a ) Kinetic profiles of the cold-induced reversible gelation of 0.25% agar at 25 °C (absorbance changes at 350 nm plotted vs. time) and their corresponding fitted curves (continuous line, ―). According to Equation (3), the fitting parameters of agar are A = 0.132, B = 12.087, C = 1.731, m = 0.00044, and q = −0.009. Derived kinetical parameters for agar gelation are Abs Max = 0.132, V Max = 0.007 Abs/min, t VMax = 5.9 min, and t AbsMax/2 = 11.9 min. Considering the crucial timings t 0 , t lag , t VMax , t AbsMax/2 and t AbsMax , five phases can be identified: I (lag phase), IIa (increase up to maximum rate), IIb (approximately constant-rate increase), IIIa (decelerating increase), and IIIb (plateau with a slight asymptotic drift); see main text for further details; ( b ) Kinetic profiles of the enzymatic assay of 8 μM S–2238 catalyzed by 0.15 nM human alpha thrombin at 25 °C (absorbance changes at 405 nm plotted vs. time) and their corresponding fitted curves (continuous line, ―). According to Equation (3), the fitting parameters for S–2238 are A = 0.139, B = 15.048, C = 1.742, m = −0.000027, and q = 0.005. Derived kinetical parameters for S–2238 enzymatic assay are Abs Max = 0.136, V Max = 0.006 Abs/min, t VMax = 7.2 min, and t AbsMax/2 = 14.9 min. Contrariwise, for the enzymatic reaction only IIa, IIb, IIIa and IIIb phases can be distinguished.

Article Snippet: Human alpha thrombin was purchased by Enzyme Research Laboratories (South Bend, IN, USA).

Techniques: Derivative Assay, Enzymatic Assay