Project description:The objective of this study was to provide insight to the molecular mechanisms chaging in the peripheral blood after acute ischemic cerebrovascular syndrome. Peripheral whole blood samples were collected from N=34 MRI diagnosed ischemic stroke patients >=18 years of age within 24 hours from known onset of symptom and again at 24-48 hours after onset. RNA was extracted from whole blood stabilized in Paxgene RNA tubes.
Project description:Acute stroke, with ischemic stroke comprising 80% of all cerebrovascular incidents, has been recognized as one of the core problems in clinical medicine in need of prevention and treatment. Intravenous rtPA is the mainstay and the highest class evidence based method of acute ischemic stroke treatment, and is currently recommended 0-4.5 hours after stroke onset. In most patients decision on i.v. rtPA administration is striaghtforward, however the biggest concern is the symptomatic intracranial haemorrhage (sICH), which occurs in 3-7% of all treated patients, and is associated with worse 90-day functional outcome and higher disability than in those untreated. Finding a method of the powerful (highly specific and selective) identification of patients at highest risk of sICH, in order to increase the percentage of stroke patients safely treated with rtPA, is one of the most important challenges in stroke research. To address this problem we designed a major and complex project to identify blood, neuroimaging, and clinical biomarkers combined for prospective assessment of the risk of intracranial hemorrhage (ICH) after thrombolytic treatment of acute ischemic stroke. Herein, we reveal our general methodological approach with shortlisting of blood protein candidates selected with Sequential Window Acquisition of All Theoretical Mass Spectra (SWATH-MS) that in the future might increase sensitivity and selectivity of the rtPA-associated sICH risk calculations.
Project description:The current treatment options for ischemic stroke aim to achieve reperfusion but are time critical. Novel therapeutic approaches that can be given beyond the limited time window of 3 - 4.5 hours are still an unmet need to be addressed to improve stroke outcome. The lack of oxygen and glucose in the area of ischemic injury initiates a pathological cascade leading to blood-brain barrier (BBB) breakdown, inflammation and neuronal cell death, a process that may be intercepted to limit stroke progression. Pericytes located at the blood/brain interface are one of the first responders to hypoxia in stroke and therefore a potential target cell for early stroke interventions. Using single-cell RNA sequencing in a mouse model of permanent middle cerebral artery occlusion, we investigated the temporal differences in transcriptomic signatures in pericytes at 1, 12, and 24 hours after stroke compared to the contralateral hemisphere. Our results reveal a stroke-specific subcluster of pericytes that is present at 12 and 24 hours and characterized by the upregulation of genes mainly related to cytokine signalling and immune response. This study identifies temporal transcriptional changes in the acute phase of ischemic stroke that reflect the early response of pericytes to the ischemic insult and its secondary consequences and may constitute potential future therapeutic targets.
Project description:Gene expression profiling of elutriated monocytes cultured for 48 hours in RPMI-1640, 10% AB plamsa and one of the following: GMCSF (56IU/ml), IL-4 (2000 IU/ml), 15 kDa Granulysin (10 nM) Sources tested: monocytes form 3 donors tested at time 0, 4 hours, 12 hours, 24 hours, 48 hours
Project description:Transcriptional profiling of MeJA-treated pearl millet seedlings over time [0, 12, 24 and 48 hours post treatment (hpt)]. Keywords: Time course, Stress response
Project description:Transcriptional profiling of SA-treated pearl millet seedlings over time [0, 12, 24 and 48 hours post treatment (hpt)]. Keywords: Time course, Stress response
Project description:The purpose of this project was to elucidate gene expression in the peripheral whole blood of acute ischemic stroke patients to identify a panel of genes for the diagnosis of acute ischemic stroke. Peripheral blood samples were collected in Paxgene Blood RNA tubes from stroke patients who were >18 years of age with MRI diagnosed ischemic stroke and controls who were non-stroke neurologically healthy. The results suggest a panel of genes can be used to diagnose ischemic stroke, and provide information about the biological pathways involved in the response to acute ischemic stroke in humans. Total RNA extracted from whole blood in n=39 ischemic stroke patients compared to n=24 healthy control subjects.
Project description:Prospective cohort studies and meta-analyses examining the relationship between HDL-cholesterol (C) and stroke risk are discordant and question the value of HDL-C as a marker for stroke risk prediction. However, the relationship between other metrics of HDL, function and proteome, and stroke remain unexplored. We investigated changes in HDL protein composition and function -cholesterol efflux capacity (CEC)- after acute ischemic stroke, and their relationship to long-term functional and neuronal recovery after stroke. Plasma samples were collected from healthy controls (N = 35) and from stroke patients either 24 (early, N = 35) or 96 hours (late, N = 20) after stroke onset. ABCA1 mediated CEC was measured using murine macrophages. Stroke recovery was assessed at 3 months after stroke event using the Modified Rankin Scores (MRS) and the NIH Stroke Scale (NIHSS). Proteomic analysis of HDL by parallel reaction monitoring indicated a distinct time-dependent remodeling after stroke. 15 proteins were significantly altered following stroke, with 6 proteins significantly changing between 24 and 96 h post stroke. Inflammation-related proteins (SAA1, SAA2, PON1) were increased at both time points. Macrophage CEC, consistent with inflammatory remodeling of HDL proteome, was reduced by 50% (P<0.0001) in both early and late post-stroke samples compared to the controls. Changes in 6 post-stroke significantly correlated with stroke recovery scores (P<0.05). Further, the multiple linear regression model adjusting for baseline stroke severity confirmed that these proteins that predict the stroke recovery. Changes in HDL associated proteins within the first 96 hours post stroke could be used as markers to predict functional stroke recovery.
Project description:This study aims to implement methods for toxicological profiling of engineered nanomaterials using toxicogenomics, proteomics, and metabolomics along with computational analyses. For all three omics layers the human cell lines A549 (lung epithelial cells) and THP1 (monocytes) were separately exposed to the nanomaterials TiO2 NM104, MWNCT NM401, and Ag NM300k. Proteomics and metabolomics samples have been performed as biological triplicates and were taken after 0, 6, 12, 24, and 48 hours. To assess ecotoxic effects C. elegans worms were grown in soil treated with NM300k. Ecotox samples were taken only at 0 and 24 hours. Integrating all three omics layers will enable the identification of (novel) ENM specific modes of action (MoA).
Project description:Ischemic stroke is a serious medical condition that leads to the onset of neurological symptoms such as loss of motor and cognitive functions. Focal cerebral ischemia (FCI) occurs due to the interrupted blood supply to the site of injury, resulting in the demise of brain cells. The subventricular zone (SVZ) is a source of stem and progenitor cells (NS/PCs), which proliferate and differentiate in multiple cell types in order to substitute the injured tissue. We isolated cells from the SVZ and the adjacent striatum three days after middle cerebral artery occlusion (MCAO) in order to map cellular changes and NS/PCs plasticity in adult brain upon injury. Expression profiling of the isolated tissue at the single cell level enables the identification of cell population subtypes in the region and their characteristic gene expression.