Als (not shown), suggesting that HT was not playing a majorAls (not shown), suggesting that

Als (not shown), suggesting that HT was not playing a major
Als (not shown), suggesting that HT was not playing a major role in their functional deterioration and early death. LDLR blockade by RAP may provide a unifying solution to a number of rt-PA modes of action which lead to BBB disruption and HT during thrombolysis [22, 23]. We therefore tested the effects of RAP against rt-PA-induced BBB breakdown and formation of HT as well as Necrosulfonamide manufacturer mortality and functional outcome in our 4 PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/25447644 h transient thread occlusion protocol, which to the best of our knowledge has not been attempted before. As observed in our initial characterisation (Fig. 1), rt-PA treatment induced a similar increase in albumin extravasation as seen for vehicle-treated mice (Fig. 2a, b) but caused a significant upsurge in the brain haemoglobin content measured 24 h after stroke (p < 0.05, Fig. 2c, d). Administration of RAP (2 mg/kg) at reperfusion with or without rt-PA did not significantly decrease total brain albumin levels (Fig. 2a), yet a trend for reduction of total brain haemoglobin was observed (p = 0.18, Fig. 2c). Normalisation of the ipsilateral value to the contralateral baseline for each mouse (i.e. fold increase analysis) revealed near-significant reductions in cerebral albumin (p = 0.054, Fig. 2b) andNiego et al. Fluids Barriers CNS (2017) 14:Page 6 ofFig. 2 Receptor-associated protein (RAP) does not significantly attenuate rt-PA-induced BBB disruption and formation of intracerebral haemorrhage. Receptor-associated protein (2 mg/kg) was co-administered with rt-PA (10 mg/kg) post 4 h middle cerebral artery occlusion (MCAo) and blood components were measured in the perfused brain 20 h later. a Albumin values in the ipsilateral hemisphere (after subtraction of the contralateral values to correct for perfusion efficiency) and b fold analysis of brain albumin (ipsilateral above contralateral, accounting for each animal's own baseline). Administration of RAP together with rt-PA does not reduce albumin levels compared to rt-PA alone (a) but a trend for reduced albumin with RAP is apparent by fold (b). c, d Brain haemoglobin is increased with rt-PA compared to vehicle both in the raw analysis (ipsilateral minus contralateral; c) or the relative analysis (fold; ipsilateral above contralateral; d). While a trend emerges, addition of RAP with rt-PA does not significantly attenuate blood levels in the brain. e Representative images of brain tissue 24 h after rt-PA or rt-PA + RAP treatment post 4 h MCAo. Significant intraparenchymal hematomas can still be observed after RAP treatment. Data is shown as individual animals with mean ?SEM. n = 11 for vehicle, 10 for rt-PA, 8 for rt-PA + RAP and 7 for RAP. One-way ANOVA with Sidak post hoc analysis of selected groups. Outliers are denoted in black symbols and excluded from the analysishaemoglobin content (p = 0.06, Fig. 2d) by RAP. Indeed, focal haematomas and diffuse bleeding across the infarct could often be observed in rt-PA + RAP-treated brains24 h post MCAo, as seen with rt-PA alone albeit to a milder extent (Fig. 2e), in line with the biochemical findings. As mentioned above, our autopsies of mice whichNiego et al. Fluids Barriers CNS (2017) 14:Page 7 ofFig. 3 Receptor-associated protein (RAP) reduces mortality and neurological score but does not improve other functional parameters. Mortality rates (a) and neurological deficit assessment (b ) 20 after 4 h middle cerebral artery occlusion following treatment with HEPES vehicle, rt-PA (10 mg/kg), RAP (2 mg/kg) or PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/28242652 their combinat.