Journal: Biochimica et biophysica acta

Article Title: Increases in intracellular calcium perturb blood-brain barrier via protein kinase C-alpha and apoptosis.

doi: 10.1016/j.bbadis.2015.10.016

Figure Lengend Snippet: Fig. 4. Amiloride and apocynin reduce PKC activity and uPA activity affected by PKC-α. HBMEC were exposed to 4 h OGD (OGD) with or without reperfusion (R) in the absence or presence of inhibitors for uPA (amiloride), NADPH oxidase (apocynin), intracellular calcium (BAPTA-AM) or PKC-α (RO-32-0432). Amiloride and apocynin decreased the rise in total PKC (A) and PKC-α (B) activities seen during OGD ± R. tPA activity is increased during OGD ± R and treatment with BAPT-AM and RO-32-0432 show no effect (C). uPA activity is also increased during OGD ± R and BAPTA-AM and RO-32-0432 normalised the increases seen (D). Data represented as mean ± SEM from n ≥3.* P b 0.05 compared to Normoxia. † P b 0.05 compared to re- spective untreated group.

Article Snippet: PAI-1-bound active uPA or tPA was detected by anti-tPA or anti-uPA primary antibodies (1:50, Santa Cruz Biotechnology) and primary antibodies were probed with horseradish peroxidase-linked secondary antibodies (1:5000, Santa Cruz).

Techniques: Activity Assay

Journal: Journal of Thrombosis and Haemostasis

Article Title: Non‐severe COVID‐19 is associated with endothelial damage and hypercoagulability despite pharmacological thromboprophylaxis

doi: 10.1111/jth.15660

Figure Lengend Snippet: Plasma tissue plasminogen activator (t‐PA) levels are significantly elevated in coronavirus disease 2019 (COVID‐19). Plasma levels of t‐PA (A) were significantly elevated in hospitalized patients with non‐severe COVID‐19 ( n = 14) compared to a group of hospitalized patients ( n = 11) who had screened negative for severe acute respiratory syndrome coronavirus 2 by real‐time polymerase chain reaction. Levels of soluble thrombomodulin (sTM; B), vascular cell adhesion molecule‐1 (VCAM‐1; C), intercellular adhesion molecule‐1 (ICAM‐1; D), plasminogen activator inhibitor‐1 (PAI‐1; E) and E‐selectin (F) were similar in both groups. All samples were assayed in technical duplicate. Data are presented as the mean ± standard deviation. Statistical analysis was performed using a two‐tailed Student’s t test. * P < .05

Article Snippet: ELISAs for TFPI (DTFP10), tissue plasminogen activator (t‐PA; DTPA00), thrombomodulin (DTHBD0), vascular cell adhesion molecule‐1 (VCAM‐1; DVC00), intercellular adhesion molecule‐1 (ICAM‐1; DCIM00), and E‐selectin (DSLE00) were purchased from R&D Systems and a plasminogen activator inhibitor‐1 (PAI‐1; ab108891) ELISA was purchased from Abcam.

Techniques: Clinical Proteomics, Real-time Polymerase Chain Reaction, Standard Deviation, Two Tailed Test

Journal: CNS neuroscience & therapeutics

Article Title: Asparagine Endopeptidase Inhibition Attenuates Tissue Plasminogen Activator-Induced Brain Hemorrhagic Transformation After Ischemic Stroke.

doi: 10.1111/cns.70345

Figure Lengend Snippet: FIGURE 3 | AEP KO ameliorates tPA-associated hemorrhagic transformation in the stroke mouse model. (A, B) Representative images of hema- toma in brain slices and quantification of hemoglobin levels. Data are presented as mean ± SEM and statistical analyses are performed using Welch test followed by Dunnett T3 multiple comparisons test were applied since the P value of Levene test < 0.05. (C) Recorded brain water content to as- sess brain edema at 24 h after delayed tPA administration. Data are presented as mean ± SEM and statistical analysis is performed using one-way ANOVA test followed by Tukey's multiple comparisons test. (D) ELISA for tPA activity detection. Data are presented as mean ± SEM and statistical analyses are performed using Welch test followed by Dunnett T3 multiple comparisons test were applied since the P value of Levene test < 0.05. (E) Representative images of HE staining in the cortical hematoma area (scale bar = 50 μm). (A), (C) n = 6, (D) n = 4 per group. Normality and variance are assessed via Shapiro-Wilk test and Levene's test, respectively. *P < 0.05, **P < 0.01.

Article Snippet: A human Tissue- type Plasminogen Activator ELISA kit (E- EL- H2106, Elabscience) was used for detection.

Techniques: Transformation Assay, Enzyme-linked Immunosorbent Assay, Activity Assay, Staining