Journal: Pharmaceuticals

Article Title: Acetonitrile Adducts of Tranexamic Acid as Sensitive Ions for Quantification at Residue Levels in Human Plasma by UHPLC-MS/MS

doi: 10.3390/ph14121205

Figure Lengend Snippet: Structure of the TXA-ACN protonated adduct formed by positive-mode electrospray ionization and the two possible protonation sites of tranexamic acid.

Article Snippet: High purity (≥99%) analytical standards of tranexamic acid (TXA, ) and the corresponding isotopically labeled internal standard 13 C 2 , 15 N , trans -tranexamic acid (TXA-IS) were supplied by Toronto Research Chemicals (Toronto, Canada).

Techniques:

Journal: Frontiers in Oncology

Article Title: Lysyl oxidase-like 4 promotes the invasiveness of triple-negative breast cancer cells by orchestrating the invasive machinery formed by annexin A2 and S100A11 on the cell surface

doi: 10.3389/fonc.2024.1371342

Figure Lengend Snippet: Annexin A2 can bind with plasminogen by fashioning a complex with S100A11. (A) Endogenous lysyl oxidase-like 4 (LOXL4), annexin A2, and S100A11 proteins in the indicated cell lines were detected using the Western blotting (WB) procedure. The running gel was also stained with Coomassie brilliant blue (CBB) as a sample control of proper loading. (B) We evaluated the invasion ability of MDA-MB-231 cells after the treatment with or without tranexamic acid (TXA) (1 mg/mL). (C) HEK293T cells were co-transfected with HA-tagged S100A11 and Myc-tagged ANXA2. After precipitation of the expressed products with the beads, bound foreign proteins were analyzed by WB using the HA or Myc antibody. (D) Prepared recombinant proteins (GST, GST-ANXA2, S100A10, and S100A11) from the Escherichia coli expression system were checked for their purity. (E) Paired blots of the indicated proteins [GST, GST-annexin A2, and S100A10 (upper) or S100A11 (lower)] were dot-blotted onto the nitrocellulose filter membranes in a duplicate manner. The blotted membranes were then incubated with either biotinylated plasminogen (bio-PLG) alone or a blend of bio-PLG and S100A10 (upper) or S100A11 (lower). After incubation was completed, the bound proteins were detected by anti-S100A10 or anti-S100A11 (left) or streptavidin–horseradish peroxidase (HRP) (right). (F) The schematic diagram resumed the results from panel (E) The red or blue circles correspond to those highlighted in the results in panel (E) Data from panel (B) are means ± SD, ***p < 0.001. The individual symbols (*)mean exploit of preparations of the purified recombinant proteins as their predicted sizes.

Article Snippet: Tranexamic acid (TXA) and MMP9 inhibitor [MMP-9-IN-1 (OUN87710)] were purchased from LKT Laboratories (St. Paul, MN, USA) and Selleck Chemicals (Houston, TX, USA), respectively.

Techniques: Western Blot, Staining, Control, Transfection, Recombinant, Expressing, Incubation, Purification

Journal: Frontiers in Oncology

Article Title: Lysyl oxidase-like 4 promotes the invasiveness of triple-negative breast cancer cells by orchestrating the invasive machinery formed by annexin A2 and S100A11 on the cell surface

doi: 10.3389/fonc.2024.1371342

Figure Lengend Snippet: Transition of plasminogen to active-form plasmin on the cell surface annexin A2/S100A11 complex. (A) Biotinylated plasminogen (bio-PLG) was evaluated for the dose used in its transition reaction to active-form plasmin in MDA-MB-231 cell culture. A treatment duration of 60 min was scheduled. To confirm the ripe plasmin band originating from the added bio-PLG, its reaction stopper tranexamic acid (TXA) (1 mg/mL) was used. (B) The temporal evaluation of the added bio-PLG was performed according to a protocol similar to that described in the former experiment (A) , except for using only 10 µg/mL. (C) The effect of calcium (Ca 2+ , 10 mM) on plasmin and other protease activities was studied. The samples prepared from the cell membrane fractions under non-reducing [dithiothreitol (DTT) (−)] conditions were subjected to gelatin zymography. After running, the gels were soaked in a refolding buffer and then incubated with a reaction buffer with (lower) or without Ca 2+ (upper). (D) Non-triple-negative breast cancer (TNBC) MCF-7 cells and the indicated TNBC cell lines were all treated with bio-PLG at a final concentration of 10 µg/mL for 60 min. The bio-PLG attached to the cell membrane and the manifestation of its active derivative in the individual cell membranes were detected by Western blotting (WB) and zymography. (E) HEK293T cells were co-transfected with HA-tagged full-length or any of several S100A11 variants lacking the C-terminal end (see Supplementary Figure 2C ) and Myc-tagged annexin A2. The transfected cells were further treated with bio-PLG (10 µg/mL, 60 min). After fractionation of the membrane compartment from each cell treated, half of the extracts were used for input detection and zymography experiments. The remaining half was subjected to a pull-down procedure using streptavidin-conjugated beads. After precipitation of the samples with the beads, bound proteins were analyzed by WB using streptavidin–horseradish peroxidase (HRP), HA antibody, or Myc antibody. (F) The indicated cells were all treated with bio-PLG (10 µg/mL, 60 min). TGF-β (10 ng/mL, 24 h) stimulation induced intrinsic lysyl oxidase-like 4 (LOXL4) production in the parental MDA-MB-231 cells. After fractionation of the membrane compartment from each cell treated, similar experiments were performed as in panel (E) except for the use of the indicated antibodies to evaluate the endogenous proteins of interest. Cont: non-engineered MDA-MB-231 cells. (G) In vitro , a cell-free reaction was performed to evaluate the effect of annexin A2/S100A11 complex on the emergence of plasmin from inactive plasminogen. The prepared, purified proteins were mixed and incubated according to the formula. After a 30-min reaction at room temperature, the samples were subjected to gelatin zymography.

Article Snippet: Tranexamic acid (TXA) and MMP9 inhibitor [MMP-9-IN-1 (OUN87710)] were purchased from LKT Laboratories (St. Paul, MN, USA) and Selleck Chemicals (Houston, TX, USA), respectively.

Techniques: Cell Culture, Membrane, Zymography, Incubation, Concentration Assay, Western Blot, Transfection, Fractionation, In Vitro, Purification

Journal: Frontiers in Oncology

Article Title: Lysyl oxidase-like 4 promotes the invasiveness of triple-negative breast cancer cells by orchestrating the invasive machinery formed by annexin A2 and S100A11 on the cell surface

doi: 10.3389/fonc.2024.1371342

Figure Lengend Snippet: Contribution of cell surface plasmin to activation of MMP9 and invasion of cancer cells. (A) A transwell-based invasion assay was performed on MDA-MB-231 cells treated with plasminogen (PLG; 10 µg/mL, 12 h) under the presence or absence of tranexamic acid (TXA) (1 or 10 mg/mL). (B) Prepared conditioned media from 1-day serum-free cultivation of the indicated cells were condensed 10-fold. The media specimens were subjected to sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and then underwent Western blotting (WB) analysis to detect MMP9 after being transferred to a polyvinylidene difluoride (PVDF) membrane or Coomassie brilliant blue (CBB) staining of the running gel without the transfer procedure, which was used as a sample control for proper loading. (C) Zymography using the gelatin substrate was performed to detect MMP-secretory activity from the indicated cells. (D) Schematic representation of the in vivo experimental lung tropic cancer metastasis protocol. MDA-MB-231 GFP cells (8 × 10 5 cells) with or without TXA (0.5 g/kg) were injected intravenously into BALB/c-nu/nu female mice. (E) Lung metastasis was monitored by images (right) from cancer-based white foci in the dissected mouse lung, and the clear foci (left) were counted as total foci (more than 1 mm in diameter). (F) Schematic representation of the molecular interplay among the indicated vital molecules. Data from panels (A, E) are means ± SD. ns: not significant, *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Tranexamic acid (TXA) and MMP9 inhibitor [MMP-9-IN-1 (OUN87710)] were purchased from LKT Laboratories (St. Paul, MN, USA) and Selleck Chemicals (Houston, TX, USA), respectively.

Techniques: Activation Assay, Transwell Invasion Assay, Polyacrylamide Gel Electrophoresis, SDS Page, Western Blot, Membrane, Staining, Control, Zymography, Activity Assay, In Vivo, Injection

Journal: Acta Orthopaedica

Article Title: Reduction of blood loss in primary hip arthroplasty with tranexamic acid or fibrin spray

doi: 10.3109/17453674.2011.623568

Figure Lengend Snippet: Patient data

Article Snippet: Patients were randomized to 1 of 3 treatment groups: (1) tranexamic acid (Cyklokapron; Meda Pharmaceuticals, UK), (2) fibrin spray (Quixil; Omrix Biopharmaceuticals, Belgium), or (3) neither (control).

Techniques: Control

Journal: The Cochrane Database of Systematic Reviews

Article Title: Drugs to reduce bleeding and transfusion in major open vascular or endovascular surgery: a systematic review and network meta‐analysis

doi: 10.1002/14651858.CD013649.pub2

Figure Lengend Snippet: Reporting of review endpoints

Article Snippet: Blinding of participants and personnel (performance bias) All outcomes , Low risk , "Patients randomised to the intervention group received tranexamic acid (Ugurol; Rottapharm S.p.A., Milan, Italy; 0.5 g 5 ml ‐1 vial) according to the following protocol: a loading dose of 500 mg of tranexamic acid diluted in 100 ml saline was intravenously infused slowly 20 min before surgery, and a continuous intravenous infusion of tranexamic acid was then administered at a rate of 250 mg h (2.5 ml h ‐1 , using non‐diluted tranexamic acid contained in the vials) from surgical incision until skin closure. Patients randomised to the control group received placebo (saline) with identical volumes and rates of infusion." "We performed a single‐centre, double‐blinded, parallel‐group, randomised clinical trial".

Techniques: Clinical Proteomics

Journal: The Cochrane Database of Systematic Reviews

Article Title: Drugs to reduce bleeding and transfusion in major open vascular or endovascular surgery: a systematic review and network meta‐analysis

doi: 10.1002/14651858.CD013649.pub2

Figure Lengend Snippet: Summary of findings: Tranexamic acid versus placebo

Article Snippet: Blinding of participants and personnel (performance bias) All outcomes , Low risk , "Patients randomised to the intervention group received tranexamic acid (Ugurol; Rottapharm S.p.A., Milan, Italy; 0.5 g 5 ml ‐1 vial) according to the following protocol: a loading dose of 500 mg of tranexamic acid diluted in 100 ml saline was intravenously infused slowly 20 min before surgery, and a continuous intravenous infusion of tranexamic acid was then administered at a rate of 250 mg h (2.5 ml h ‐1 , using non‐diluted tranexamic acid contained in the vials) from surgical incision until skin closure. Patients randomised to the control group received placebo (saline) with identical volumes and rates of infusion." "We performed a single‐centre, double‐blinded, parallel‐group, randomised clinical trial".

Techniques: Comparison, Control

Journal: The Cochrane Database of Systematic Reviews

Article Title: Drugs to reduce bleeding and transfusion in major open vascular or endovascular surgery: a systematic review and network meta‐analysis

doi: 10.1002/14651858.CD013649.pub2

Figure Lengend Snippet: Transfusion outcomes for trials of systemic drugs

Article Snippet: Blinding of participants and personnel (performance bias) All outcomes , Low risk , "Patients randomised to the intervention group received tranexamic acid (Ugurol; Rottapharm S.p.A., Milan, Italy; 0.5 g 5 ml ‐1 vial) according to the following protocol: a loading dose of 500 mg of tranexamic acid diluted in 100 ml saline was intravenously infused slowly 20 min before surgery, and a continuous intravenous infusion of tranexamic acid was then administered at a rate of 250 mg h (2.5 ml h ‐1 , using non‐diluted tranexamic acid contained in the vials) from surgical incision until skin closure. Patients randomised to the control group received placebo (saline) with identical volumes and rates of infusion." "We performed a single‐centre, double‐blinded, parallel‐group, randomised clinical trial".

Techniques: Clinical Proteomics

Journal: The Cochrane Database of Systematic Reviews

Article Title: Drugs to reduce bleeding and transfusion in major open vascular or endovascular surgery: a systematic review and network meta‐analysis

doi: 10.1002/14651858.CD013649.pub2

Figure Lengend Snippet: Systemic drug treatments

Article Snippet: Blinding of participants and personnel (performance bias) All outcomes , Low risk , "Patients randomised to the intervention group received tranexamic acid (Ugurol; Rottapharm S.p.A., Milan, Italy; 0.5 g 5 ml ‐1 vial) according to the following protocol: a loading dose of 500 mg of tranexamic acid diluted in 100 ml saline was intravenously infused slowly 20 min before surgery, and a continuous intravenous infusion of tranexamic acid was then administered at a rate of 250 mg h (2.5 ml h ‐1 , using non‐diluted tranexamic acid contained in the vials) from surgical incision until skin closure. Patients randomised to the control group received placebo (saline) with identical volumes and rates of infusion." "We performed a single‐centre, double‐blinded, parallel‐group, randomised clinical trial".

Techniques: Comparison

Journal: The Cochrane Database of Systematic Reviews

Article Title: Drugs to reduce bleeding and transfusion in major open vascular or endovascular surgery: a systematic review and network meta‐analysis

doi: 10.1002/14651858.CD013649.pub2

Figure Lengend Snippet: Monaco 2020

Article Snippet: Blinding of participants and personnel (performance bias) All outcomes , Low risk , "Patients randomised to the intervention group received tranexamic acid (Ugurol; Rottapharm S.p.A., Milan, Italy; 0.5 g 5 ml ‐1 vial) according to the following protocol: a loading dose of 500 mg of tranexamic acid diluted in 100 ml saline was intravenously infused slowly 20 min before surgery, and a continuous intravenous infusion of tranexamic acid was then administered at a rate of 250 mg h (2.5 ml h ‐1 , using non‐diluted tranexamic acid contained in the vials) from surgical incision until skin closure. Patients randomised to the control group received placebo (saline) with identical volumes and rates of infusion." "We performed a single‐centre, double‐blinded, parallel‐group, randomised clinical trial".

Techniques: Molecular Weight, Filtration, Clinical Proteomics, Sequencing, Selection, Blocking Assay, Generated, Saline, Control

Journal: The Cochrane Database of Systematic Reviews

Article Title: Drugs to reduce bleeding and transfusion in major open vascular or endovascular surgery: a systematic review and network meta‐analysis

doi: 10.1002/14651858.CD013649.pub2

Figure Lengend Snippet: POISE‐3 2022

Article Snippet: Blinding of participants and personnel (performance bias) All outcomes , Low risk , "Patients randomised to the intervention group received tranexamic acid (Ugurol; Rottapharm S.p.A., Milan, Italy; 0.5 g 5 ml ‐1 vial) according to the following protocol: a loading dose of 500 mg of tranexamic acid diluted in 100 ml saline was intravenously infused slowly 20 min before surgery, and a continuous intravenous infusion of tranexamic acid was then administered at a rate of 250 mg h (2.5 ml h ‐1 , using non‐diluted tranexamic acid contained in the vials) from surgical incision until skin closure. Patients randomised to the control group received placebo (saline) with identical volumes and rates of infusion." "We performed a single‐centre, double‐blinded, parallel‐group, randomised clinical trial".

Techniques: Saline, Clinical Proteomics, Sequencing, Selection, Blocking Assay

Journal: The Cochrane Database of Systematic Reviews

Article Title: Drugs to reduce bleeding and transfusion in major open vascular or endovascular surgery: a systematic review and network meta‐analysis

doi: 10.1002/14651858.CD013649.pub2

Figure Lengend Snippet: jRCTs041180163

Article Snippet: Blinding of participants and personnel (performance bias) All outcomes , Low risk , "Patients randomised to the intervention group received tranexamic acid (Ugurol; Rottapharm S.p.A., Milan, Italy; 0.5 g 5 ml ‐1 vial) according to the following protocol: a loading dose of 500 mg of tranexamic acid diluted in 100 ml saline was intravenously infused slowly 20 min before surgery, and a continuous intravenous infusion of tranexamic acid was then administered at a rate of 250 mg h (2.5 ml h ‐1 , using non‐diluted tranexamic acid contained in the vials) from surgical incision until skin closure. Patients randomised to the control group received placebo (saline) with identical volumes and rates of infusion." "We performed a single‐centre, double‐blinded, parallel‐group, randomised clinical trial".

Techniques: Control

Journal: The Cochrane Database of Systematic Reviews

Article Title: Drugs to reduce bleeding and transfusion in major open vascular or endovascular surgery: a systematic review and network meta‐analysis

doi: 10.1002/14651858.CD013649.pub2

Figure Lengend Snippet: NCT04803747

Article Snippet: Blinding of participants and personnel (performance bias) All outcomes , Low risk , "Patients randomised to the intervention group received tranexamic acid (Ugurol; Rottapharm S.p.A., Milan, Italy; 0.5 g 5 ml ‐1 vial) according to the following protocol: a loading dose of 500 mg of tranexamic acid diluted in 100 ml saline was intravenously infused slowly 20 min before surgery, and a continuous intravenous infusion of tranexamic acid was then administered at a rate of 250 mg h (2.5 ml h ‐1 , using non‐diluted tranexamic acid contained in the vials) from surgical incision until skin closure. Patients randomised to the control group received placebo (saline) with identical volumes and rates of infusion." "We performed a single‐centre, double‐blinded, parallel‐group, randomised clinical trial".

Techniques: Saline, Biomarker Discovery