MSJ741191_supplementary_appendix_a – Supplemental material for Fulminant rebound of relapsing–remitting multiple sclerosis after discontinuation of dimethyl fumarate: A case report

<p>Supplemental material, MSJ741191_supplementary_appendix_a for Fulminant rebound of relapsing–remitting multiple sclerosis after discontinuation of dimethyl fumarate: A case report by Peter Harmel, Frieder Schlunk and Lutz Harms in Multiple Sclerosis Journal</p

MSJ741191_supplementary_appendix_b – Supplemental material for Fulminant rebound of relapsing–remitting multiple sclerosis after discontinuation of dimethyl fumarate: A case report

<p>Supplemental material, MSJ741191_supplementary_appendix_b for Fulminant rebound of relapsing–remitting multiple sclerosis after discontinuation of dimethyl fumarate: A case report by Peter Harmel, Frieder Schlunk and Lutz Harms in Multiple Sclerosis Journal</p

Gelatin zymography of MMP responses.

<p>(<b>A</b>) Representative zymogram gels showing MMP-2 and MMP-9 levels in brain homogenates derived from mice subjected to focal cerebral ischemia (induced by middle cerebral artery occlusion, MCAO) or primary ICH mice. (<b>B</b>) Quantified densitometry of the zymogram results (arbitrary units normalized to constant loading of MMP standards). After 3 hrs of focal cerebral ischemia followed by 3 hrs reperfusion, tPA significantly amplified MMP-2 and MMP-9 levels. In contrast, there were no differences in MMPs in saline versus tPA-treated mice after ICH.</p

Collagenase-induced ICH volumes are not increased by administration of i.v. tPA.

<p>(<b>A</b>) Brain sections showing hematoma at 30 min, 60 min and 24 hrs after ICH induction. ICH starts to occur within 30 min and has largely developed by 1 hr. (<b>B</b>) Hematoma volumes at 24 hrs after ICH induction (mean±SD). tPA did not alter hematoma sizes, but heparin significantly worsened ICH as compared to saline and tPA treatment. (<b>C</b>) Results of the reconfirmation study. No difference was observed between saline and tPA mice, but heparin significantly increased ICH volume (mean±SD) as compared to saline and tPA treatment.</p

Treatment with tPA worsened subarachnoid hemorrhage.

<p>Blood volumes at 24 hrs after SAH induction (mean±SEM) are provided. tPA significantly increased SAH blood volume (tPA, n = 6; saline, n = 7).</p

tPA activity measures.

<p>(<b>A</b>) Human tPA but not saline activates mouse plasminogen. In this assay, tPA converts plasminogen to plasmin, then plasmin converts D-Val-Leu-Lys-7-Amino-4-Methylcoumarin to a fluorescent product, which was measured over 1 hr at 5 minute intervals using a fluorescence plate reader. Data are provided in arbitrary fluorescence units (A.U.). Data are mean±SEM, 3 measures per group and time point. (<b>B</b>) Thrombolytic activity of tPA was proven in-vivo in a standard model of thromboembolic middle cerebral artery occlusion in rats. tPA restored regional cerebral blood flow (rCBF) more rapidly than saline. Data are mean±SEM, 4 animals per group.</p