We want to relegate the key generator functionality to an external web server so that we can use our local server some something entirely different. So essentially, I have the algorithm written in a proprietary language, and I need to choose a cPanel supported environment on our external webserver to re-implement the algorithm in such a way that it can be called by the client's browser, updating a database, without the client being able to download the subroutine and view the code.
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You need an additional Ansible tarball, which contains Ansible and other dependent packages. Download ansible.tar.gz and store it anywhere outside the installer folder. For download instructions, see ansible.tar.gz.If ansible.tar.gz is located in some different location, update the absolute location for the --offline-prerequisites-bundle parameter.
SCMECs were seeded on a Matrigel basement membrane matrix (BD, CA, USA) to examine tube formation. Matrigel (50μ l) was added to the center of each well of a 96-well plate. The plate was then allowed to solidify for 30min at 37C. After exposure to OGD and incubation with STS for 24h, 1.5104 cells/well were plated to form tubes. Cells were allowed to incubate for an appropriate period of time at 37C (5% CO2). Photographs were acquired using an inverted microscope (200x). The number of junctions and the total length of tubes were counted using ImageJ.
A total of 30 adult male C57BL/6 mice (20-25g) were randomly divided into 3 groups (n=10 per group) including the sham-operated, SCI model, and STS-treated groups. Allen's method was used to establish the SCI model with a moderate contusion, based on previous methods [32]. Mice were anesthetized using pentobarbital sodium (30mg/kg, i.p.). Spinal cords were exposed through T9-T10 laminectomy under sterile conditions followed by a contusion with an impact velocity of 0.5m/s, depth of 0.6mm, and duration of 80ms. Successful establishment of the SCI model was observed by spinal cord congestion, leg swaying, tail swing reflexes, and slow paralysis. Next, the bladder was manually pressed twice a day. STS (dissolve in saline, 20mg/kg) was administered through intraperitoneal injection at 2h after SCI once a day for a week. Similar methodology was applied to the vehicle groups. After treatment, mice were sacrificed using pentobarbital sodium (80mg/kg, i.p.), and spinal cords (5mm sections of the spinal cord centered at the lesion site) were extracted and stored at -80C for immunofluorescence and western blotting experiments or were perfused with paraformaldehyde for histopathological evaluation (H&E) and Nissl staining. The detailed experimental design for this study is shown in supplementary materials (Figure 1S).
Figure 2: Morphological characterization and immunofluorescence analysis of SCMECs. (a) Representative fields of SCMEC morphologies at the primary, first, and third passages (scale bar=200μ m, 100μ m, and 50μ m). (b) Representative immunofluorescent labeling images for CD31, vWF, NG-2, and GFAP (scale bar=50μ m) [Please download the PDF to view the image]. 2ff7e9595c
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