11 research outputs found

    Effect of VPA on the secondary structure of rhPE.

    No full text
    <p>(<b>A</b>) Three VPA concentrations (6, 12 and 24 mM) were titrated into 2.5 µM rhPE. VPA caused a significant change in the secondary structure of the enzyme. (<b>B</b>) A high concentration of ZPP (1 µM) was added to the 2.5 µM rhPE/24 mM VPA mixture. ZPP did not cause any change in the secondary structure of the enzyme. (<b>C</b>) Adding 1 µM ZPP to 2.5 µM rhPE does not cause any secondary protein structure change. Adding 24 mM VPA on top of that mixture doesn't cause any structure change.</p

    VPA decreases cigarette smoke-induced neutrophil influx in BAL fluid of mice.

    No full text
    <p>(<b>A</b>) Total cell numbers, (<b>B</b>) macrophages and (<b>C</b>) neutrophils in the BAL fluid of mice exposed to air or whole body cigarette smoke twice daily during 5 days. The mice received vehicle (PBS) or VPA (100 µg/70 µl PBS) by oropharyngeal aspiration 15 minutes prior to air/smoke exposure. (<b>D</b>) acPGP was measure in the BAL fluid. (<b>E</b>) PE activity was measured in the BAL fluid and compared to control (PBS treated/air exposed mice). N = 5–10 animals per group. Values are expressed as mean+/−S.E.M. **P≤0.01, ***P≤0.001, ns p>0.05.</p

    NMR spectra of the carbobenzoxy-group of ZPP.

    No full text
    <p>(<b>A</b>) 10 µM ZPP was added to 10 µM rhPE, 10 mM VPA was added to that mixture. A shift of the free ZPP peaks at 7.28 and 7.20 ppm to the left is seen (indicated in the red and green bars respectively). (<b>B</b>) 10 µM ZPP was added to 10 µM rhPE. A shift of the free ZPP peaks at 7.28 and 7.20 ppm to the left is seen. (<b>C</b>) 10 µM ZPP was measured without enzyme. The peaks at 7.28 and 7.20 ppm are the free ZPP fractions. (<b>D</b>) 10 mM VPA was added to 10 µM rhPE, 10 µM ZPP was added to that mixture. Note that the free ZPP peaks at 7.28 and 7.20 ppm do not show a shift to the left; there are no peaks in the highlighted areas.</p

    Inhibition of PGP generation by VPA.

    No full text
    <p>(<b>A</b>) Dialyzed Collagen Type I and II were incubated with the lysate of 4.6 * 10<sup>6</sup> PMN at 37°C for 20 hours to generate PGP. (<b>B</b>) PE activity was measured 30 minutes after incubation of lysate/collagen/VPA mixture and compared to control (no VPA). Data are shown as the mean ± S.E.M. (n = 5–10 per group). Representative of 4 experiments. # p<0.0001 compared to control, $ p<0.01 compared to control, ** p<0.01, ns p>0.05.</p

    Effect of VPA on the activity of rhPE.

    No full text
    <p>(<b>A</b>) Activity of 10 nM purified rhPE was measured in presence of ten doses of VPA or lithium ranging from 0.2–10 mM. Relative activity of rhPE in the presence of VPA or lithium is shown as a percentage of activity in the absence of VPA and lithium. VPA showed a <i>K<sub>i</sub></i> of approximately 1 mM. lithium showed no effect on PE activity. (<b>B</b>) Inhibition curves of three VPA concentrations (0.8, 1.6 and 3.5 mM) were obtained by incubating VPA with 10 nM rhPE during 90 min at 37 °0. (<b>C</b>) A Lineweaver-Burk plot was made based on the rhPE activity assays with 0.8 and 1.6 mM VPA as inhibitor. Enzyme activity was measured with increasing substrate (Suc-Gly-Pro-pNA) concentrations, ranging from 0.2–10 mM. The velocity was calculated as mM * min<sup>−1</sup> * ml<sup>−1</sup>. Data are shown as the mean ± S.E.M. (n = 3 per group).</p

    Immunostaining for phosphorylated Focal Adhesion Kinase.

    No full text
    <p>MSCs were seeded onto glass coverslips coated with electrospun nanofibers, or with FBS as a control. After 5 hours, cells were fixed and stained for phosphorylated Focal Adhesion Kinase (red). Cells were counterstained with DAPI to show cell nuclei (blue). Cells seeded onto PCL/col/HA scaffolds were better spread, and exhibited greater amounts of punctuate pFAK staining (site pY397) as compared with cells on PCL or PCL/HA. Cells seeded onto FBS-coated glass coverslips displayed pFAK staining in focal adhesion-type structures (white arrows), as expected for cells grown on 2D surfaces.</p

    Live cell imaging of GFP-expressing MSCs seeded onto electrospun scaffolds.

    No full text
    <p>A) Cells were seeded onto scaffolds and imaged over varying time points. Panels a–c: PCL scaffolds; panels d–f: PCL/HA scaffolds; panels g–i: PCL/col/HA scaffolds and panels j–l: col scaffolds. Scale bar = 100 µm. B) Higher magnification images of GFP-expressing MSCs at seven hours on electrospun scaffolds (panels m–p).</p

    Adsorption of FN and VN by electrospun scaffolds.

    No full text
    <p>Scaffolds were coated with fetal bovine serum (A), or implanted into rat tibial osteotomies for 30 min (B). Scaffolds were then washed to remove loosely bound proteins, and proteins were subsequently desorbed by incubation in boiling SDS-containing solution. The amounts of FN and VN were evaluated by Western blot.</p

    Tensile Properties of dry and hydrated scaffolds.

    No full text
    <p>Values represent the average ± standard deviation calculated in the linear portion at 10% strain. The hydrated collagen scaffolds have very low mechanical properties and could not be measured by this technique.</p

    MTS assay quantifying cell proliferation on electrospun scaffolds of PCL, PCL/HA or PCL/col/HA.

    No full text
    <p>At day one, cell number was significantly higher on PCL/HA and PCL/col/HA scaffolds in comparison to PCL. By day four, PCL/HA was still significantly higher than PCL, and PCL/col/HA was significantly higher than PCL/HA and PCL. In addition, cell number on PCL/col/HA was significantly higher on day four than day one. An * denotes p<0.05</p
    corecore