Legislative Documents

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Flow cytometry (FC) analysis of whole blood samples from healthy controls (H), non-thrombocytopenic septic (Non-T) and septic thrombocytopenic (T) donors.

<p><b>A–D</b>, Non-significant differences in the platelet surface expression of MMP-2, -9, TLR-4 and P-selectin between the 3 groups represented as the mean plus the standard error of the mean. <b>E</b>, Gating of the platelet population based on characteristic side and forward scatter. <b>F</b>, Representative graph from FC analysis in a thrombocytopenic septic patient demonstrating the presence of MMP-9 on platelets. The signal intensity of the MMP-9 antibody is highlighted by the red circle. In this case 85.7% of the gated platelet population is positive for the presence of MMP-9. <b>G</b>, Response of healthy platelets to agonist TRAP-6 with increased expression of TLR-4 and P-selectin. <b>H</b>, Response of platelets from septic patients to TRAP-6 with increased expression only in P-selectin. Data are mean ± SEM. n = 8 healthy controls, n = 5–9 non-thrombocytopenic donors, n = 3–5 thrombocytopenic donors. *p < 0.05 and **p < 0.01.</p

Examination of the activity of MMP-2 and -9 by gelatin zymography in septic shock as a 3 compartment model.

<p>Results are shown firstly as both healthy controls (H) and septic patients (S) and then with the septic patients categorised as non-thrombocytopenic (Non-T) and thrombocytopenic (T) patients. <b>A and B</b>, PPP samples at protein concentration (100μg/lane)–H (n = 20), Non-T (n = 13) and T (n = 7). <b>C and D</b>, Intraplatelet or lysate samples at protein concentration 30μg/lane–H (n = 10), Non-T (n = 7) and T (n = 5). <b>E and F</b>, Homogenate or plasma membrane samples at maximal protein concentration of 7.9 μg/lane–H (n = 9), Non-T (n = 5) and T (n = 5). <b>G</b>, Representative gel zymography showing both healthy controls (lane 1 and 2) and septic donors (lane 3); control = lane 4 (HT1080 media). <b>H</b>, Zymography gels. <b>1</b> shows PPP sample on the left lane and positive control on the right lane. On the left lane, a small band can be seen directly under the upper line. This small line corresponds to the active MMP-9 (82 kDa). This line is not visible in <b>2</b>, where lysate samples of septic patients were evaluated. Data are mean ± SEM. Results are compared to healthy controls *p < 0.05, **p < 0.01 and ***p < 0.001.</p

Exclusion criteria for patient selection.

<p>Exclusion criteria for patient selection.</p

Survival outcomes.

<p>Plasma levels of MMP-9 were significantly higher in non-survivors (n = 6) compared to survivors (n = 14) (p = 0.005). No association was found between MMP-2 levels in PPP and mortality (p = 0.477).</p

PLA formation in sepsis.

<p><b>A</b>, Increased PLA formation in the thrombocytopenic (T) group (n = 4) compared to the non-thrombocytopenic (Non-T) group (n = 9); healthy (H) controls (n = 8). <b>B</b>, Immunohistochemical staining of a large PLA from a septic thrombocytopenic donor using a 10x objective lens magnification (100x). In the enhanced picture the large red arrows point to leukocytes and the smaller blue arrows point to platelets. Leukocytes are larger, nucleated and stained violet with haematoxylin. Platelets are smaller, stained with anti-CD62P and are rust in colour.</p