Dade actin fsl reagent

CBT test was conducted using 15 rats (3 from each experimental group). The toenail of the animals’ hind paws was cut where the nail met the quick, using a single-edged razor blade, before being immediately immersed in Ringer’s solution at 37 °C. Maximum bleeding was defined as the time after the operator used external pressure to halt the bleeding [55 (link)]. While observing with a binocular microscope, with 3-times magnification, the time until the cessation of the bleeding (without rebleeding within 30 s) was calculated.
Clotting time measures, such as PT and APTT, were evaluated to identify the effect of the drug-loaded nanoformulations on the coagulation cascade in contrast to the free APX suspension and control groups [56 (link),57 (link)]. Blood samples from 3 rats (for each study group) were obtained 120 min after drug administration. The plasma was obtained as previously described in the PK section, before then being immediately utilized to assess PT and APTT. The Dade Thromboplastin-C reagent, obtained from the Baxter Healthcare Corporation (Deerfield, IL, USA) was utilized to assess PT using the technique of mechanical clot detection with an International Sensitivity Index (ISI) of 2.0. By contrast, APTT was assessed using both the mechanical clot detection technique and the Siemens Healthineers Dade^® Actin^® FSL reagent (Erlangen, Germany) [58 (link)].

Blood samples were collected into 2.7 mL vacuum tubes containing 3.2% sodium citrate (Becton-Dickinson Company, New Jersey, USA) for aPTT analysis. The samples were centrifuged (Andreas Hettich GmbH & Co. KG, Tuttlingen, Germany) at 1500xg for 15 minutes. Platelet-poor plasma (PPP) was obtained with a verified platelet count of less than 10 x10⁹/L. Dade Actin FSL reagent (Siemens Healthcare, Marburg, Germany) containing purified soy phosphatides and rabbit brain phosphatides in 1.0 x 10^-4M ellagic acid was used for the determination of aPTT within one hour of collection. Activated partial thromboplastin time was analysed using the Sysmex CS2100i automated coagulation analyser (Sysmex Corporation, Kobe, Japan). Samples with haematocrit exceeding 55% and haemolysed samples were not included in the analysis. The assay variation in our laboratory is maintained below 5%.

aPTT-based CWA data were retrieved from the CS2100i automated coagulation analyser (Sysmex Corporation, Kobe, Japan). The Dade Actin FSL reagent (Siemens Healthcare, Marburg, Germany) used for the determination of aPTT contains purified soy phosphatides and rabbit brain phosphatides in 1.0 × 10^–4 M ellagic acid. A built-in algorithm tool on the analyzer generates the clotting curve measured at 660 nm and three other CWA parameters – maximum velocity (min1), maximum acceleration (min2), maximum deceleration (max2) and the corresponding timings (Tmin1, Tmin2, Tmax2)^{17 (link)}. The parameters used for comparison between the groups were the subjects’ aPTT, min1, min2 and max2 values.