Involvement of Matrix Metalloproteinase in Thrombolysis-Associated Hemorrhagic Transformation After Embolic Focal Ischemia in Rats
|
|
- Sheryl Justina Neal
- 5 years ago
- Views:
Transcription
1 Involvement of Matrix Metalloproteinase in Thrombolysis-Associated Hemorrhagic Transformation After Embolic Focal Ischemia in Rats Toshihisa Sumii, MD, PhD; Eng H. Lo, PhD Background and Purpose Thrombolytic therapy with tissue plasminogen activator (tpa) for acute ischemic stroke remains complicated by risks of hemorrhagic transformation. In this study we used a previously established quantitative rat model of tpa-associated hemorrhage to test the hypothesis that matrix metalloproteinases (MMPs) are involved. Methods Spontaneously hypertensive rats were subjected to embolic focal ischemia by placing homologous blood clots into the middle cerebral artery. Three groups of rats were studied: (1) untreated controls that received saline at 6 hours after ischemia; (2) rats that received tpa alone (10 mg/kg at 6 hours after ischemia); and (3) rats that received tpa plus the broad-spectrum MMP inhibitor BB-94 (50 mg/kg of BB-94 before ischemia and at 3 and 6 hours after ischemia plus tpa at 6 hours). Gelatin zymography was used to quantify MMP levels. A hemoglobin spectrophotometry method was used to quantify cerebral hemorrhage. Ischemic lesions were measured at 24 hours with tetrazolium staining. Results At 6, 12, and 24 hours, pro-mmp-9 and cleaved MMP-9 were upregulated in ischemic brain. At 12 hours, tpa-treated rats showed significantly higher levels of pro-mmp-9 and cleaved MMP-9 than untreated controls. By 24 hours, all rats showed evidence of hemorrhagic transformation in the ischemic territory. Rats treated with BB-94 and tpa showed significantly reduced hemorrhage volumes compared with those that received tpa alone. There was no effect on infarct size. Conclusions These results indicate that (1) tpa treatment increases levels of MMP-9 after embolic focal cerebral ischemia, (2) MMPs are involved in the mechanism of tpa-associated hemorrhage, and (3) combination therapies with MMP inhibitors may be useful for decreasing the risk and severity of this dreaded complication of thrombolytic therapy. (Stroke. 2002;33: ) Key Words: cerebral hemorrhage extracellular matrix reperfusion injury stroke tissue plasminogen activator rats Although thrombolytic therapy with tissue plasminogen activator (tpa) may be effective for acute ischemic stroke, 1,2 there is an elevated risk of cerebral hemorrhage. 3,4 In part, nonspecific damage to cerebrovascular walls due to free radical generation during reperfusion injury has been implicated. 5,6 However, the precise molecular mechanisms that underlie this dreaded complication remain to be fully elucidated. Recently, there has been an emphasis on the possible role of proteases that are upregulated after cerebral ischemia and reperfusion. Specifically, the class of zinc-dependent matrix metalloproteinases (MMPs) has been intensively investigated. In mouse, rat, and baboon models of cerebral ischemia, expression of several MMPs is significantly increased after ischemic onset MMPs can degrade almost all components of extracellular matrix, and therefore uncontrolled activation of these enzymes can result in significant tissue damage. In the context of hemorrhagic transformation after cerebral ischemia, MMPs may degrade vascular basal lamina, weaken vessels, and predispose them to rupture. In experimental studies, activation of MMP-9 and degradation of critical protein components of cerebral blood vessels have been correlated with the development of hemorrhage and edema. 8,12,17 In a recent study, pharmacological inhibition of MMPs significantly decreased the incidence of hemorrhage in a rabbit model of embolic stroke. 18 However, it remained unclear whether there were any effects on the severity of hemorrhage. Surprisingly, a qualitative assessment of hemorrhage size suggested that it may have actually been worsened by MMP inhibitor treatment. If MMP inhibition is to be validated as a therapeutic approach against tpa-associated Received August 23, 2001; final revision received October 31, 2001; accepted December 6, From the Neuroprotection Research Laboratory, Departments of Neurology and Radiology, Massachusetts General Hospital, Charlestown (T.S., E.H.L.); Program in Neuroscience, Harvard Medical School, Boston, Mass (E.H.L.); and Department of Neurosurgery, Kinki University School of Medicine, Osaka-Sayama, Japan (T.S.). Correspondence to Eng H. Lo, PhD, Neuroprotection Research Laboratory, Departments of Neurology and Radiology, Harvard Medical School, MGH East , Charlestown, MA Lo@helix.mgh.harvard.edu 2002 American Heart Association, Inc. Stroke is available at 831
2 832 Stroke March 2002 hemorrhagic transformation, it will be important to carefully assess its effects not only on risks but on severity of hemorrhage as well. We have previously characterized a quantitative model of tpa-associated cerebral hemorrhage in rat embolic stroke. 19 In the present study we use this model to assess the effects of tpa on the profiles of MMP-2 and MMP-9 upregulation and the efficacy of the broad-spectrum MMP inhibitor BB-94 (batimastat) for reducing the volume of hemorrhage. Materials and Methods Embolic Model of Focal Cerebral Ischemia All experiments were performed following an institutionally approved protocol in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals. Male spontaneously hypertensive rats (Taconic, Germantown, NY) were used. Animals were anesthetized with halothane (1% to 1.2%) under spontaneous respiration in a 30% O 2 /70% N 2 O mixture. Rectal temperatures were maintained at C with a thermostatcontrolled heating pad. The right femoral artery was cannulated, and physiological parameters, including rectal temperature, mean arterial blood pressure, ph, PCO 2, and PO 2, were monitored throughout all experiments. The right femoral vein was cannulated for drug administration. Focal ischemia was induced with the use of homologous blood clots following methods that have been previously described Briefly, femoral arterial blood from a donor rat was withdrawn into 50 cm of polyethylene tubing (PE-50), kept in the tube for 2 hours at room temperature, and subsequently retained for 22 hours at 4 C. Five centimeters of the PE-50 tubing containing the clot was cut and connected to a syringe filled with saline with a 23-gauge needle. The clot was transferred into a dish filled with saline and washed 5 times with a 30-cm PE-10 tube. Then the clot was shifted to a modified PE-50 catheter with a 0.3-mm outer diameter filled with saline. Under a surgical microscope (Carl Zeiss, Inc), a modified PE-50 catheter with a 5-cm-long blood clot was gently inserted into the external carotid artery until the tip was positioned just proximal to the origin of middle cerebral artery. Then the clot in the catheter was injected into the internal carotid artery along with small amount of saline. After 5 minutes, the catheter was withdrawn from the external carotid artery. Reperfusion was achieved with the use of tpa. Continuous laser-doppler flowmetry (Perimed AB) was used to monitor regional cerebral perfusion to ensure adequacy of embolic occlusions (perfusion decreased to 15% of preischemic baselines). For placement of the laser-doppler flowmetry probe, a burr hole 2 to 3 mm in diameter was created in the right parietal bone (2 mm posterior and 6 mm lateral to bregma). Experimental Groups and Drug Treatments The goals of this study were (1) to assess the profiles of MMP upregulation after rat embolic stroke with and without tpa treatment and (2) to assess the effects of the broad-spectrum MMP inhibitor BB-94 on tpa-associated hemorrhage. Three groups of rats were studied: (1) untreated controls that received saline at 6 hours after ischemia; (2) rats that received tpa alone (10 mg/kg at 6 hours after ischemia); and (3) tpa plus BB-94 treated rats that received 50 mg/kg of BB-94 before ischemia and at 3 and 6 hours after ischemia plus tpa at 6 hours. tpa (Activase, Genentech) was administered intravenously (10 mg/kg, 2 mg/ml concentration in saline, over 20 minutes) according to previously established protocols that have been demonstrated to effectively induce clot lysis and cerebral reperfusion in rat embolic stroke The relatively high dose of tpa was chosen on the basis of the approximately 10-fold difference in fibrin-specific activity between human and rodent systems. 26 BB-94 was dissolved in PBS with 0.01% Tween-80 (3 mg/ml) and given intraperitoneally. Timing for the 3 doses of BB-94 was based on the estimated 3-hour plasma half-life of BB-94 in rodents. 27 Similar doses have been shown to be effective inhibitors of MMP activity in rodent tumor models. 28 Furthermore, the intraperitoneal route of BB-94 administration has been previously demonstrated to effectively penetrate brain, 29 and we have previously shown that this dose of BB-94 worked in a mouse model of focal cerebral ischemia. 7 BB-94 was a kind gift from British Biotech. SDS-PAGE Gelatin Zymography Gelatin zymography was used to measure the levels of MMP-2 and MMP-9 following previously described techniques. 7,8,30 One set of rats was used to document the time course of MMP upregulation at 6, 12, and 24 hours after onset of embolic stroke (n 3 rats per group). Another set of rats was used to compare the profiles of ischemic upregulation in MMP between untreated rats and those treated with tpa. These measurements were performed at 12 and 24 hours (n 5 per group). In all cases, rats were killed after transcardial perfusion with ice-cold PBS (ph 7.4). The brains were quickly removed, divided into ipsilateral ischemic hemispheres and contralateral nonischemic hemispheres, then frozen immediately in liquid nitrogen and stored at 80 C to await further procedures. The brain samples were homogenized in 10 volume of lysis buffer (50 mmol/l Tris-HCl [ph 7.4] containing 150 mmol/l NaCl, 1% Igepal CA-630, 0.1% SDS, and 0.1% deoxycholic acid) including protease inhibitors (2 g/ml leupeptin, 2 g/ml aprotinin, and 1 mmol/l phenylmethylsulfonyl fluoride) on ice with the use of a Teflon glass homogenizer. After centrifugation ( rpm) for 7 minutes at 4 C, the supernatant fluid was collected. The total protein concentration of each sample was determined by the Bradford assay (Bio-Rad Laboratories). Equal volumes of total protein extracts normalized for protein concentration (15 g/5 L) were prepared. Then 5 L of 2 sample buffer (126 mmol/l Tris-HCl, 20% glycerol, 4% SDS, 0.005% bromophenol blue) was added to each protein extract. Protein samples were loaded on and separated by 10% Tris-glycine gel with 0.1% gelatin as substrate. After separation by electrophoresis, the gel was incubated in renaturing buffer (2.7% Triton X-100 in distilled water) at room temperature for 30 minutes with gentle agitation. Then the renaturing buffer was decanted and replaced with developing buffer (50 mmol/l Tris base, 40 mmol/l HCl, 200 mmol/l NaCl, 5 mmol/l CaCl 2, 0.2% Brij 35). After 30 minutes of equilibration by the developing buffer, the gel was incubated with fresh developing buffer at 37 C for 24 hours. After developing, the gel was stained with 0.5% Coomassie blue R-250 for 30 minutes and then destained appropriately. Human MMP-2 and MMP-9 standards were purchased from Chemicon. For densitometry, gels were scanned and analyzed with a standard computerassisted imaging system. Elevations in MMP were calculated as fold increase in optical density versus nonischemic normal brain. 9,31 Analysis of Infarct Volumes and Neurological Deficits At 24 hours after ischemia, rats were assessed with a 4-point neurological deficit scale that has been extensively used for rat models of stroke. 32 After neurological assessment, rats were killed with a lethal overdose of sodium pentobarbital and transcardially perfused to remove all intravascular blood. Coronal brain sections (2 mm thick) were stained with triphenyltetrazolium chloride (TTC) (Sigma). Infarct volumes were quantified according to the indirect method 33 via standard computer-assisted image analysis techniques. Spectrophotometric Assay of Intracerebral Hemorrhage Cerebral hemorrhage was quantified with a previously described spectrophotometric assay. 19,34 Initially, a standard curve was obtained with the use of a virtual model of hemorrhage. Hemispheric brain tissue was obtained from normal rats subjected to complete transcardial perfusion to remove intravascular blood. Incremental volumes of homologous blood (0, 0.5, 1, 2, 4, 8, 16, 32, 50, 100, 200 L) were added to each hemispheric sample with PBS to reach a total volume of 3 ml, followed by homogenization for 30 seconds, sonication on ice for 1 minute, and centrifugation at rpm for 30 minutes. Drabkin s reagent (1.6 ml; Sigma) was added to 0.4-mL
3 Sumii and Lo MMPs and tpa-associated Hemorrhage 833 Figure 1. Gelatin zymogram showing upregulation of MMP-9 after embolic focal cerebral ischemia in spontaneously hypertensive rats. Both pro-mmp-9 and cleaved MMP-9 levels increased over time. Slight elevation in MMP-9 was observed in the contralateral hemisphere as well. Ips indicates ipsilateral brain; cont, contralateral brain; and P.C., standards of human MMP-2 and MMP-9. Normal nonischemic brain is designated as control. aliquots and allowed to stand for 15 minutes at room temperature. Optical density was measured and recorded at 540 nm with a spectrophotometer (Spectronix 3000, Milton-Roy). These procedures yielded a linear relationship between hemoglobin concentrations in perfused brain and the volume of added blood. Measurements from perfused brains subjected to ischemia were compared with this standard curve to obtain data in terms of hemorrhage volume ( L). Hemorrhage measurements were performed on brains already stained with TTC for infarct quantitation. We have previously established that TTC staining does not alter the spectrophotometric hemoglobin assay. 19 Statistical Analysis Quantitative densitometry data from the zymograms were analyzed by ANOVA followed by Tukey s honestly significant difference tests. Hemorrhage and infarct volumes were compared by 2-tailed t tests. Neurological deficits were compared by Mann-Whitney U tests. Mortality rates were compared by the 2 test. Data were expressed as mean SEM. Probability values 0.05 were considered significant. Results Normal rat brain showed detectable baseline levels of MMP-2 but not MMP-9 (Figure 1). After injection of homologous blood clots to induce focal embolic ischemia, MMP-9 levels increased over time. Both pro-mmp-9 (92 kda) and cleaved MMP-9 (84 kda) were observed (Figure 1). Interestingly, the contralateral hemisphere also showed signs of increased MMP-9 levels (Figure 1). To assess the effects of tpa treatment on these ischemic profiles of MMP upregulation, untreated rats were compared with rats treated with tpa (administered at 6 hours after ischemia). In all rats MMP-9 levels were elevated, as expected. At 12 hours, tpa-treated rats showed significantly higher levels of pro-mmp-9 and cleaved MMP-9 than untreated rats (Figure 2A to 2C). No significant differences were noted for MMP-2 (Figure 2A and 2D). By 24 hours after ischemia, overall MMP levels increased, but there were no longer any significant differences between untreated rats and tpa-treated rats (Figure 2A to 2D). A separate series of experiments was performed to assess the effects of MMP inhibition on tpa-associated hemorrhage in this rat model of embolic stroke. Ischemic rats treated with tpa alone were compared with rats treated with tpa plus BB-94. All rats showed physiological parameters within normal range (Table 1). As expected, mean arterial blood pressures were high for these spontaneously hypertensive Figure 2. A, Representative gelatin zymogram showing the effects of tpa on the ischemic upregulation in MMP-9 at 12 and 24 hours. tpa-treated rats showed higher levels of pro-mmp-9 and cleaved MMP-9 than untreated rats at 12 hours after focal ischemia. Standard densitometry techniques were used to quantify these changes as fold increase versus nonischemic baselines (mean SEM) for pro-mmp-9 (B), cleaved MMP-9 (C), and pro-mmp-2 (D). *P 0.05; n 5 per group for each time point. P.C. indicates standards of human MMP-2 and MMP-9. rats. Injection of homologous blood clots resulted in immediate and uniform reductions in cerebral perfusion, as documented by laser-doppler flowmetry (Table 2). Delayed tpa treatment (6 hours after ischemia) in this model resulted in a mortality rate of 50% (7 of 14 rats). Rats treated with a combination of tpa plus BB-94 had a significantly reduced mortality rate (9.1%; 1 of 11 rats) compared with controls that received tpa alone (P 0.01) (Table 3). In surviving rats (n 7 tpa only; n 10 tpa plus BB-94), hemorrhagic transformation, ischemic lesion volumes, and neurological deficits were assessed. Spectrophotometric measurement of whole blood showed a linear response between blood volume and hemoglobin absorbance (Figure 3A), thus validating this method for quantifying hemorrhage. By 24 hours after ischemia, all rats showed evidence of hemorrhagic transformation within the ischemic zone, demonstrating the reproducibility of this model of tpa-associated hemorrhage. Treatment with BB-94 significantly reduced hemorrhage volumes by almost 50% compared with vehicle-treated controls (Figure 3B). There were no effects on ischemic lesion volumes (Figure 3C) or neurological deficits (Table 3). Discussion Thrombolysis with tpa can elevate risks of cerebral hemorrhage after ischemic stroke. 3,4 Thus, it is important to dissect
4 834 Stroke March 2002 TABLE 1. Physiological Variables Group Weight, g Rectal Temperature, C MABP, mm Hg ph PCO 2, mm Hg PO 2, mm Hg Vehicle (n 7) BB-94 (n 10) MABP indicates mean arterial blood pressure. Values are mean SEM. the mechanisms involved and identify methods that can ameliorate hemorrhage risks after tpa stroke therapy. In this report we used a previously established quantitative model of hemorrhage in spontaneously hypertensive rats to show that (1) pro-mmp-9 and cleaved MMP-9 are upregulated after focal ischemia induced by a blood clot embolus, (2) tpa treatment further increased the levels of pro-mmp-9 and cleaved MMP-9, and (3) combination therapy with the broad-spectrum MMP inhibitor BB-94 significantly reduced the volume of tpa-associated cerebral hemorrhage. Hemorrhagic transformation after tpa thrombolysis may be broadly related to deleterious events during reperfusion injury. Historically, reperfusion injury has been associated with the generation of oxygen-derived free radicals. 35 Free radicals damage proteins and lipids in cell membranes, thus amplifying cell death. 36,37 In the context of hemorrhage, it is conceivable that free radical damage to membranes of the cerebrovascular system during reperfusion injury would lead to vascular leakage or rupture. In rodent models of embolic focal ischemia induced with homologous blood clots, delayed treatment with tpa has been observed to result in hemorrhagic transformations Experimental interventions using free radical scavengers effectively reduce the severity of hemorrhage after tpa-induced reperfusion. 19,38,39 These data provide hope that combination therapies that target free radical mechanisms can be designed to ameliorate risks of hemorrhagic transformation after thrombolytic reperfusion. In addition to free radical mediated reperfusion injury, another emerging candidate mechanism may involve MMPs. It has been shown that MMPs are upregulated after acute central nervous system injury after ischemia, 11 13,15 hemorrhage, 40 and trauma. 30,41 43 As a family of extracellular proteases, MMPs can degrade almost all components of the extracellular matrix. In the context of hemorrhagic transformation, uncontrolled MMP activation after tpa reperfusion can degrade critical proteins in the cerebrovasculature. These include collagen and laminin in the basal lamina 12,17 and the blood-brain barrier associated protein ZO-1. 8 Degradation of these critical components may disrupt vascular structural integrity and lead to leakage and rupture. Interestingly, it has been shown that free radical injury is mechanistically linked to MMP upregulation. 42 Hence, the involvement of MMPs in TABLE 2. Regional Cerebral Blood Flow Group Immediately After Ischemia, % 10 min After Ischemia, % Vehicle (n 7) BB-94 (n 10) Values are mean SEM. the pathophysiology of hemorrhage fits well in the context of oxidative damage and reperfusion injury. Recent data also suggest that tpa is biochemically linked to the MMP axis of extracellular proteolysis, with plasmin acting as an upstream activator of the MMP cascade. 44,45 This provides another potential mechanism for the role of MMP-mediated hemorrhage after tpa thrombolysis. It will be critical for future studies to carefully examine the mechanisms and effects of tpa on all potential sources of MMP activity, including inflammatory cells that may infiltrate through the disrupted blood-brain barrier. In the present study we showed that pro-mmp-9 and cleaved MMP-9 were both upregulated after embolic focal ischemia in rats. Importantly, delayed treatment with tpa significantly enhanced this increase in pro-mmp-9 and cleaved MMP-9. These data suggest that MMP-9 may be involved in the process of tpa-associated hemorrhage. A recent study demonstrated that cotreatment with the MMP inhibitor BB-94 reduced the incidence of hemorrhage in a rabbit model of embolic stroke. 18 However, the effect of MMP inhibition on the severity of hemorrhage was unclear; the data suggested that the MMP inhibitor may have actually increased the severity of hemorrhage when it occurred. For clinical applications, it will be important to assess the effects not only on risk or incidence but also on severity or volume of hemorrhage. We have previously established a quantitative model of tpa-associated cerebral hemorrhage after clot-based embolic focal ischemia in spontaneously hypertensive rats. 19 Clinically, the relevance of this model may be related both to the presence of hypertension and the concomitant vascular phenotype within this strain of rats. Nevertheless, the incidence of tpa-associated hemorrhage in this model is 100%, and therefore this reproducible model allows us to test the effects of BB-94 on the severity or volume of hemorrhage. In the present study we showed that BB-94 reduced the volume of tpa-associated hemorrhage by almost 50%. These findings implicate a role for MMPs in tpa-associated cerebral hemorrhage and suggest that targeting the MMP cascade may be a useful therapeutic approach. There are a few caveats in this study worth discussing. First, we administered BB-94 before and during ischemia. Our purpose here was to obtain proof that MMPs were involved in mediating the occurrence of hemorrhagic transformation after tpa. For clinical purposes, it will be important to determine whether delayed treatment after tpa administration will also reduce hemorrhage volume. Second, we did not directly assay for BB-94 effects on brain MMP activity. However, others have previously shown that this class of hydroxymate-based inhibitors can indeed decrease zymographic MMP activity in brain inflammation. 46 Third, it was
5 Sumii and Lo MMPs and tpa-associated Hemorrhage 835 TABLE 3. Neurological Deficits and Mortality Neurological Deficit Score Group 2 h After Ischemia 24 h After Ischemia Mortality at 24 h (Dead Animals/Surgical Animals) Vehicle (n 7) % (7/14)* BB-94 (n 10) % (1/11) Values are mean SEM. *P interesting to note that BB-94 had no effects on ischemic lesion volumes. This stands in contrast to our previous study, in which BB-94 reduced infarct volumes in a mouse model of focal ischemia. 7 However, the previous study used a mechanical method of arterial occlusion, whereas a clot-based embolic approach was used here. There may be significant differences in neuroprotective responses between mechanical versus embolic models of focal ischemia, 47,48 and further studies are needed to resolve these issues. Fourth, despite the reduction in hemorrhage volumes, we did not detect any Figure 3. A, The linear relationship between blood volume and hemoglobin absorbance in the spectrophotometric assay allowed this to be used as a standard curve for calculating hemorrhage volumes in tpa-treated brains. B, tpa-associated hemorrhage volumes (mean SEM) were significantly reduced by cotreatment with BB-94 (*P 0.05). C, Infarct volumes (mean SEM) were not significantly different in tpa only vs tpa plus BB-94 groups. improvements in neurological deficits using a relatively simple scoring system in rats. It will be useful for future studies to use more sensitive behavioral tests to define possible benefits in functional outcomes. Fifth, we used BB-94, a broad-spectrum MMP inhibitor, to ameliorate hemorrhage severity. It will be critical to ultimately define the specific MMP members and pathways involved. For example, we have shown that MMP-9 7,8 but not MMP-2 9 may be important for amplifying tissue infarction after ischemia in mouse brain. Targeting specific MMPs may lead to more optimal outcomes for preventing tpa-associated hemorrhage. Finally, in this model tpa was administered in a delayed fashion, ie, 6 hours after ischemic onset. Admittedly, tpa would not be given in such a delayed time frame in clinical stroke. Our purpose here, however, was to use a model in which hemorrhagic transformation occurred reproducibly so that we could study the mechanisms involved. The effects of MMP inhibition on the much lower rates of hemorrhage after early tpa therapy will have to be determined carefully. In conclusion, this study showed that ischemic upregulation in MMP-9 is significantly enhanced by tpa treatment after embolic stroke, and cotreatment with the broadspectrum MMP inhibitor BB-94 significantly reduced tpaassociated cerebral hemorrhage. Further studies are warranted to dissect specific MMP pathways involved, assess therapeutic windows, and translate these findings into clinical applications. Acknowledgments This study was funded by National Institutes of Health grants R01-NS37074, R01-NS38731, and R01-NS The authors would like to express their gratitude to Dr Michael Chopp for sharing his methodology for rodent embolic focal ischemia models and to Drs Pak Chan, Gary Rosenberg, and Gregory del Zoppo for very helpful advice and discussions. BB-94 was a kind donation from British Biotech. References 1. ECASS Study Group. Intravenous thrombolysis with recombinant tissue plasminogen activator in acute hemispheric stroke. JAMA. 1995;274: NINDS rt-pa Stroke Study Group. Tissue plasminogen activator for acute ischemic stroke. N Engl J Med. 1995;333: Larrue V, von Kummer R, del Zoppo G, Bluhmki E. Hemorrhagic transformation in acute ischemic stroke: potential contributing factors in the ECASS study. Stroke. 1999;28: NINDS rt-pa Stroke Study Group. Intracerebral hemorrhage after intravenous TPA therapy for ischemic stroke. Stroke. 1997;28: del Zoppo GJ, Zeumer H, Harker LA. Thrombolytic therapy in stroke: possibilities and hazards. Stroke. 1986;7: Lyden PD, Zivin JA. Hemorrhagic transformation after cerebral ischemia: mechanisms and incidence. Cerebrovasc Brain Metab Rev. 1993;5: Asahi M, Asahi K, Jung JC, del Zoppo GJ, Fini ME, Lo EH. Role of matrix metalloproteinase 9 in focal cerebral ischemia: effects of gene
6 836 Stroke March 2002 knockout and enzyme inhibition with BB-94. J Cereb Blood Flow Metab. 2000;20: Asahi M, Wang X, Mori T, Sumii T, Jung J, Moskowitz MA, Fini ME, Lo EH. Effects of matrix metalloproteinase 9 gene knockout on the proteolysis of blood-brain barrier and white matter components after cerebral ischemia. J Neurosci. 2001;21: Asahi M, Sumii T, Fini ME, Itohara S, Lo EH. Matrix metalloproteinase 2 gene knockout has no effect on acute brain injury after focal ischemia. Neuroreport. 2001;12: Fujimura M, Gasche Y, Morita-Fujimura Y, Massengale J, Kawase M, Chan PH. Early appearance of activated matrix metalloproteinase 9 and blood-brain barrier disruption in mice after focal cerebral ischemia and reperfusion. Brain Res. 1999;842: Gasche Y, Fujimura Y, Copin J, Kawase M, Masengale J, Chan PH. Early appearance of activated MMP-9 after focal cerebral ischemia in mice. J Cereb Blood Flow Metab. 1999;19: Heo JH, Lucero J, Abumiya T, Koizol JA, Copeland BR, del Zoppo GJ. Matrix metalloproteinases increase very early during experimental focal cerebral ischemia. J Cereb Blood Flow Metab. 1999;19: Mun-Bryce S, Rosenberg GA. Matrix metalloproteinases in cerebrovascular disease. J Cereb Blood Flow Metab. 1998;18: Romanic AM, White RF, Arleth AJ, Ohlstein EH, Barone FC. Matrix metalloproteinase expression increases after cerebral focal ischemia in rats. Stroke. 1998;29: Rosenberg GA, Navratil M, Barone F, Feuerstein G. Proteolytic cascade enzymes increase in focal cerebral ischemia in rat. J Cereb Blood Flow Metab. 1996;16: Rosenberg GA, Cunningham LA, Wallace J, Aleander S, Estrada EY, Grossetete M, Razhagi A, Miller K, Gearing A. Immunohistochemistry of matrix metalloproteinases in reperfusion injury to rat brain: activation of MMP-9 linked to stromelysin-1 and microglia in cell cultures. Brain Res. 2001;893: Hamman GF, Okada Y, del Zoppo GJ. Hemorrhagic transformation and microvascular integrity during focal cerebral ischemia-reperfusion. J Cereb Blood Flow Metab. 1996;16: Lapchak PA, Chapman DF, Zivin JA. Metalloproteinase inhibition reduces thrombolytic (tissue plasminogen activator)-induced hemorrhage after thromboembolic stroke. Stroke. 2000;31: Asahi M, Asahi K, Wang X, Lo EH. Reduction of tissue plasminogen activator induced hemorrhage and brain injury by free radical spin trapping after focal cerebral ischemia in rats. J Cereb Blood Flow Metab. 2000;20: Dijkhuizen RM, Asahi M, Wu O, Rosen BR, Lo EH. Delayed rt-pa treatment in a rat embolic stroke model: diagnosis and prognosis of ischemic injury and hemorrhagic transformation with magnetic resonance imaging. J Cereb Blood Flow Metab. 2001;21: Zhang ZG, Zhang RL, Jiang Q, Raman SB, Cantwell L, Chopp M. A new rat model of thrombotic focal cerebral ischemia. J Cereb Blood Flow Metab. 1997;17: Brinker G, Pillekamp F, Hossman KA. Brain hemorrhage after rtpa treatment of embolic stroke in spontaneously hypertensive rats. Neuroreport. 1999;10: Chopp M, Zhang RL, Zhang ZG, Jiang Q. The clot thickens: thrombolysis and combination therapies. Acta Neurochir Suppl (Wien). 1999;73: Kano T, Katayama Y, Tejima E, Lo EH. Hemorrhagic transformation after fibrinolytic therapy with tissue plasminogen activator in a rat thromboembolic model of stroke. Brain Res. 2000;854: Meng W, Wang X, Asahi M, Kano T, Asahi K, Ackerman RH, Lo EH. Effects of tissue type plasminogen activator in embolic versus mechanical models of focal cerebral ischemia in rats. J Cereb Blood Flow Metab. 1999;19: Korninger C, Collen D. Studies on specific fibrinolytic effect of human extrinsic plasminogen activator in human blood and in various animal species in vitro. Thromb Haemost. 1981;46: Low JA, Johnson MD, Bone EA, Dickson RB. The matrix metalloproteinase inhibitor BB-94 retards human breast cancer tumor growth but not ascites formation in nude mice. Clin Cancer Res. 1996;2: Goss KJH, Brown PD, Matrisian LM. Differing effects of endogenous and synthetic inhibitors of metalloproteinases on intestinal tumorigenesis. Int J Cancer. 1998;78: Paez Pereda M, Ledda MF, Goldberg V, Chervin A, Carrizo G, Molina H, Muller A, Renner U, Podhajcer O, Arzt E, Stalla GK. High levels of matrix metalloproteinases regulate proliferation and hormone secretion in pituitary cells. J Clin Endocrinol Metab. 2000;85: Wang X, Jung J, Asahi M, Chwang W, Russo L, Moskowitz MA, Dixon CE, Fini ME, Lo EH. Effects of matrix metalloproteinase 9 gene knockout on morphological and motor outcomes after traumatic brain injury. J Neurosci. 2000;20: Rosenberg GA, Estrada EY, Dencoff JE. Matrix metalloproteinases and TIMPs are associated with blood-brain barrier opening after reperfusion in rat brain. Stroke. 1998;29: Bederson JB, Pitts LH, Tsuji M, Nishimura MC, Davis RL, Bartkowski H. Rat middle cerebral artery occlusion: evaluation of the model and development of a neurologic examination. Stroke. 1986;17: Lin TN, Yong YH, Wa G, Khan M, Hsu CY. Effect of brain edema on infarct volume in a focal ischemia model in rats. Stroke. 1993;24: Choudri TF, Hoh BL, Solomon RA, Connoly ES, Pinsky D. Use of a spectrophotometric assay to objectively quantify intracerebral hemorrhage in mice. Stroke. 1997;28: Hallenback JM, Dutka AJ. Current concepts of reperfusion injury. Arch Neurol. 1990;47: Chan PH, Fishman RA, Longar S, Chen S, Yu A. Cellular and molecular effects of polyunsaturated fatty acids in brain ischemia and injury. Prog Brain Res. 1984;63: Chan PH. Role of oxidants in ischemic brain damage. Stroke. 1996;27: Hu B, Liu C, Zivin JA. Reduction of intracerebral hemorrhaging in a rabbit embolic stroke model. Neurology. 1999;53: Lapchak PA, Chapman DF, Zivin JA. Pharmacological effects of the spin trap agents N-t-butyl-phenylnitrone (PBN) and 2,2,6,6- tetramethylpiperidine-n-oxyl (TEMPO) in a rabbit thromboembolic stroke model: combination studies with the thrombolytic tissue plasminogen activator. Stroke. 2001;32: Rosenberg GA, Navratil M. Metalloproteinase inhibition blocks edema in intracerebral hemorrhage in the rat. Neurology. 1997;48: Ferguson TA, Muir D. MMP-2 and MMP-9 increase the neuritepromoting potential of Schwann cell basal laminae and are upregulated in degenerated nerve. Mol Cell Neurosci. 2000;16: Morita-Fujimura Y, Fujimura M, Gasche Y, Copin J, Chan PH. Overexpression of copper and zinc superoxide dismutase in transgenic mice prevents the induction and activation of matrix metalloproteinases after cold injury induced brain trauma. J Cereb Blood Flow Metab. 1999;20: Vecil GG, Larsen PH, Corley SM, Herx LM, Besson A, Goodyer CG, Yong VW. Interleukin-1 is a key regulator of matrix metalloproteinase-9 expression in human neurons in culture and following mouse brain trauma in vivo. J Neurosci Res. 2000;61: Cuzner ML, Opdenakker G. Plasminogen activators and matrix metalloproteinases, mediators of extracellular proteolysis in inflammatory demyelination of the central nervous system. J Neuroimmunol. 1999;94: Hartung HP, Kieseier BC. The role of matrix metalloproteinases in autoimmune damage to the central and peripheral nervous system. J Neuroimmunol. 2000;107: Mun-Bryce S, Rosenberg GA. Gelatinase B modulates selective opening of the blood-brain barrier during inflammation. Am J Physiol. 1998;274: R1203 R Hara T, Mies G, Hossmann KA. Effect of thrombolysis on the dynamics of infarct evolution after clot embolism of middle cerebral artery in mice. J Cereb Blood Flow Metab. 2000;20: Hata R, Maeda K, Hermann D, Mies G, Hossmann KA. Evolution of brain infarction after transient focal cerebral ischemia in mice. J Cereb Blood Flow Metab. 2000;20:
Imaging ischemic strokes: Correlating radiological findings with the pathophysiological evolution of an infarct
Imaging ischemic strokes: Correlating radiological findings with the pathophysiological evolution of an infarct Jay Chyung,, PhD, HMS III Patient A: history 91 y.o. woman Acute onset R sided weakness and
More informationAntiactin-Targeted Immunoliposomes Ameliorate Tissue Plasminogen Activator-Induced Hemorrhage After Focal Embolic Stroke
Journal of Cerebral Blood Flow & Metabolism 23:895 899 2003 The International Society for Cerebral Blood Flow and Metabolism Published by Lippincott Williams & Wilkins, Inc., Baltimore Antiactin-Targeted
More informationSUPPLEMENTARY INFORMATION
SUPPLEMENTARY INFORMATION Supplementary Figure 1. Long-term protection studies. 45 minutes of ischemia was induced in wild type (S1pr2 +/+ ) and S1pr2 -/- by MCAO. A) 5 days later brains were harvested
More informationCritical Role for Copper/Zinc Superoxide Dismutase in Preventing Spontaneous Intracerebral Hemorrhage During Acute and Chronic Hypertension in Mice
Critical Role for Copper/Zinc Superoxide Dismutase in Preventing Spontaneous Intracerebral Hemorrhage During Acute and Chronic Hypertension in Mice Yoshinobu Wakisaka, MD, PhD; Yi Chu, PhD; Jordan D. Miller,
More informationMINOCYCLINE VASCULAR PROTECTION AFTER ACUTE ISCHEMIC STROKE LIVIA SAMPAIO MACHADO. (Under the Direction of Susan C.
MINOCYCLINE VASCULAR PROTECTION AFTER ACUTE ISCHEMIC STROKE By LIVIA SAMPAIO MACHADO (Under the Direction of Susan C. Fagan) ABSTRACT Stroke is the third leading cause of death and the leading cause of
More informationInterendothelial tight junctions, the basal lamina, and
Mild to Moderate Hypothermia Prevents Microvascular Basal Lamina Antigen Loss in Experimental Focal Cerebral Ischemia Gerhard F. Hamann, MD; Dorothe Burggraf, PhD; Helge K. Martens, MD; Martin Liebetrau,
More informationAstrocytic induction of matrix metalloproteinase-9 and edema in brain hemorrhage
& 2007 ISCBFM All rights reserved 0271-678X/07 $30.00 www.jcbfm.com Astrocytic induction of matrix metalloproteinase-9 and edema in brain hemorrhage Emiri Tejima 1, Bing-Qiao Zhao 1, Kiyoshi Tsuji 1, Anna
More informationCombination Studies With the Thrombolytic Tissue Plasminogen Activator
Neuroprotective Effects of the Spin Trap Agent Disodium-[(tert-butylimino)methyl]benzene-1,3-disulfonate N-Oxide (Generic NXY-059) in a Rabbit Small Clot Embolic Stroke Model Combination Studies With the
More informationDental Research Institute, Faculty of Dentistry, University of Toronto, Toronto, Canada *For correspondence:
Zymogram Assay for the Detection of Peptidoglycan Hydrolases in Streptococcus mutans Delphine Dufour and Céline M. Lévesque * Dental Research Institute, Faculty of Dentistry, University of Toronto, Toronto,
More informationExperimental Assessment of Infarct Lesion Growth in Mice using Time-Resolved T2* MR Image Sequences
Experimental Assessment of Infarct Lesion Growth in Mice using Time-Resolved T2* MR Image Sequences Nils Daniel Forkert 1, Dennis Säring 1, Andrea Eisenbeis 2, Frank Leypoldt 3, Jens Fiehler 2, Heinz Handels
More informationHuman Urokinase / PLAU / UPA ELISA Pair Set
Human Urokinase / PLAU / UPA ELISA Pair Set Catalog Number : SEK10815 To achieve the best assay results, this manual must be read carefully before using this product and the assay is run as summarized
More informationCerebrovascular Disease lll. Acute Ischemic Stroke. Use of Intravenous Alteplace in Acute Ischemic Stroke Louis R Caplan MD
Cerebrovascular Disease lll. Acute Ischemic Stroke Use of Intravenous Alteplace in Acute Ischemic Stroke Louis R Caplan MD Thrombolysis was abandoned as a stroke treatment in the 1960s due to an unacceptable
More informationLower doses of isoflurane treatment has no beneficial effects in a rat model of intracerebral hemorrhage
Lower doses of isoflurane treatment has no beneficial effects in a rat model of intracerebral hemorrhage The Harvard community has made this article openly available. Please share how this access benefits
More informationIncreased Brain Expression of Matrix Metalloproteinase-9 After Ischemic and Hemorrhagic Human Stroke
Increased Brain Expression of Matrix Metalloproteinase-9 After Ischemic and Hemorrhagic Human Stroke Anna Rosell, PhD; Arantxa Ortega-Aznar, MD; José Alvarez-Sabín, MD, PhD; Israel Fernández-Cadenas, MSc;
More informationPrediction of Hemorrhage in Acute Ischemic Stroke Using Permeability MR Imaging
AJNR Am J Neuroradiol 26:2213 2217, October 2005 Technical Note Prediction of Hemorrhage in Acute Ischemic Stroke Using Permeability MR Imaging Andrea Kassner, Timothy Roberts, Keri Taylor, Frank Silver,
More informationChromatin Immunoprecipitation (ChIPs) Protocol (Mirmira Lab)
Chromatin Immunoprecipitation (ChIPs) Protocol (Mirmira Lab) Updated 12/3/02 Reagents: ChIP sonication Buffer (1% Triton X-100, 0.1% Deoxycholate, 50 mm Tris 8.1, 150 mm NaCl, 5 mm EDTA): 10 ml 10 % Triton
More informationSELECTIVE INHIBITION OF MATRIX METALLOPROTEINASES BY NEW HYDROXAMATE COMPOUNDS
ORIGINAL ARTICLES SELECTIVE INHIBITION OF MATRIX METALLOPROTEINASES BY NEW HYDROXAMATE COMPOUNDS ELENA GANEA 1*, CRISTINA LASLO 1, IOANA GOLDNER 1, GABRIELA PUTINA 2 1 Institute of Biochemistry of the
More informationENDOVASCULAR THERAPIES FOR ACUTE STROKE
ENDOVASCULAR THERAPIES FOR ACUTE STROKE Cerebral Arteriogram Cerebral Anatomy Cerebral Anatomy Brain Imaging Acute Ischemic Stroke (AIS) Therapy Main goal is to restore blood flow and improve perfusion
More informationSupplementary Figure 1. H-PGDS deficiency does not affect GI tract functions and anaphylactic reaction. (a) Representative pictures of H&E-stained
1 2 3 4 5 6 7 8 9 10 11 Supplementary Figure 1. H-PGDS deficiency does not affect GI tract functions and anaphylactic reaction. (a) Representative pictures of H&E-stained jejunum sections ( 200 magnification;
More informationWestern Immunoblotting Preparation of Samples:
Western Immunoblotting Preparation of Samples: Total Protein Extraction from Culture Cells: Take off the medium Wash culture with 1 x PBS 1 ml hot Cell-lysis Solution into T75 flask Scrap out the cells
More informationmm Distance (mm)
b a Magnet Illumination Coverslips MPs Objective 2575 µm 1875 µm 1575 µm 1075 µm 875 µm 545 µm 20µm 2 3 0.5 0.3mm 1 1000 100 10 1 0.1 1000 100 10 1 0.1 Field Induction (Gauss) 1.5 0 5 10 15 20 Distance
More informationProtocol for Gene Transfection & Western Blotting
The schedule and the manual of basic techniques for cell culture Advanced Protocol for Gene Transfection & Western Blotting Schedule Day 1 26/07/2008 Transfection Day 3 28/07/2008 Cell lysis Immunoprecipitation
More informationPAF Acetylhydrolase Assay Kit
PAF Acetylhydrolase Assay Kit Catalog Number KA1354 96 assays Version: 04 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Background... 3 Principle of the Assay... 3 General
More informationSUPPLEMENTARY MATERIAL
SUPPLEMENTARY MATERIAL Purification and biochemical properties of SDS-stable low molecular weight alkaline serine protease from Citrullus Colocynthis Muhammad Bashir Khan, 1,3 Hidayatullah khan, 2 Muhammad
More informationStroke Clinical Trials Update Transitioning to an Anatomic Diagnosis in Ischemic Stroke
Stroke Clinical Trials Update Transitioning to an Anatomic Diagnosis in Ischemic Stroke Alexander A. Khalessi MD MS Director of Endovascular Neurosurgery Surgical Director of NeuroCritical Care University
More informationSTUDIES ON MUSTARD-STIMULATED PROTEASES AND INHIBITORS IN HUMAN EPIDERMAL KERATINOCYTES (HEK): DEVELOPMENT OF ANTIVESICANT DRUGS
STUDIES ON MUSTARD-STIMULATED PROTEASES AND INHIBITORS IN HUMAN EPIDERMAL KERATINOCYTES (HEK): DEVELOPMENT OF ANTIVESICANT DRUGS Xiannu Jin 1, Radharaman Ray 2, Guang Xu 1 and Prabhati Ray 1 1 Department
More information7 TI - Epidemiology of intracerebral hemorrhage.
1 TI - Multiple postoperative intracerebral haematomas remote from the site of craniotomy. AU - Rapana A, et al. SO - Br J Neurosurg. 1998 Aug;1():-8. Review. IDS - PMID: 1000 UI: 991958 TI - Cerebral
More informationProtocol for protein SDS PAGE and Transfer
Protocol for protein SDS PAGE and Transfer According to Laemmli, (1970) Alaa El -Din Hamid Sayed, Alaa_h254@yahoo.com Serum Selection of a protein source cell cultures (bacteria, yeast, mammalian, etc.)
More informationProcine sphingomyelin ELISA Kit
Procine sphingomyelin ELISA Kit For the quantitative in vitro determination of Procine sphingomyelin concentrations in serum - plasma - celiac fluid - tissue homogenate - body fluid FOR LABORATORY RESEARCH
More informationThe Schedule and the Manual of Basic Techniques for Cell Culture
The Schedule and the Manual of Basic Techniques for Cell Culture 1 Materials Calcium Phosphate Transfection Kit: Invitrogen Cat.No.K2780-01 Falcon tube (Cat No.35-2054:12 x 75 mm, 5 ml tube) Cell: 293
More informationBuilding a Stroke Portfolio. June 28, 2018
Building a Stroke Portfolio June 28, 2018 1 Forward-Looking Statements This presentation contains forward-looking statements, including statements relating to: the potential benefits, safety and efficacy
More informationStroke 101. Maine Cardiovascular Health Summit. Eileen Hawkins, RN, MSN, CNRN Pen Bay Stroke Program Coordinator November 7, 2013
Stroke 101 Maine Cardiovascular Health Summit Eileen Hawkins, RN, MSN, CNRN Pen Bay Stroke Program Coordinator November 7, 2013 Stroke Statistics Definition of stroke Risk factors Warning signs Treatment
More informationDisclosures. Anesthesia for Endovascular Treatment of Acute Ischemic Stroke. Acute Ischemic Stroke. Acute Stroke = Medical Emergency!
Disclosures Anesthesia for Endovascular Treatment of Acute Ischemic Stroke I have nothing to disclose. Chanhung Lee MD, PhD Associate Professor Anesthesia and perioperative Care Acute Ischemic Stroke 780,000
More informationIschemic stroke frequently results from occlusion of cerebral
Inhibition of Factor Xa Reduces Ischemic Brain Damage After Thromboembolic Stroke in Rats Xinkang Wang, PhD; Lin Xu, MD; Hugh Wang, PhD; Reinhard Grzanna, PhD; Yutian Zhan, MS; Robert M. Knabb, PhD; Joseph
More informationSCIRF Award #2016 I-03 PI: Azizul Haque, PhD Grant Title: Neuron-specific Enolase and SCI
SCIRF Award #2016 I-03 PI: Azizul Haque, PhD Grant Title: Neuron-specific Enolase and SCI 10-month Technical Progress Report Enolase is a multifunctional glycolytic enzyme involved in growth control, hypoxia,
More informationSupplementary material: Materials and suppliers
Supplementary material: Materials and suppliers Electrophoresis consumables including tris-glycine, acrylamide, SDS buffer and Coomassie Brilliant Blue G-2 dye (CBB) were purchased from Ameresco (Solon,
More informationCerebrovascular occlusion by blood clots represents an
Mouse Model of Microembolic Stroke and Reperfusion D.N. Atochin, MD, PhD; J.C. Murciano, PhD; Y. Gürsoy-Özdemir, MD, PhD; T. Krasik; F. Noda; C. Ayata, MD; A.K. Dunn, PhD; M.A. Moskowitz, MD; P.L. Huang,
More informationAdministration of recombinant human tissue plasminogen
Effects of a Selective CD11b/CD18 Antagonist and Recombinant Human Tissue Plasminogen Activator Treatment Alone and in Combination in a Rat Embolic Model of Stroke Li Zhang, MD; Zheng Gang Zhang, MD, PhD;
More informationCollagenase Assay Kit
Collagenase Assay Kit Catalog # 31 and 32 For Research Use Only - Not Human or Therapeutic Use INTRODUCTION Collagenases are members of the matrix metalloproteinase (MMP) family and degrade collagen types
More informationACUTE STROKE TREATMENT IN LARGE NIHSS PATIENTS. Justin Nolte, MD Assistant Profession Marshall University School of Medicine
ACUTE STROKE TREATMENT IN LARGE NIHSS PATIENTS Justin Nolte, MD Assistant Profession Marshall University School of Medicine History of Presenting Illness 64 yo wf with PMHx of COPD, HTN, HLP who was in
More informationProcaspase-3. Cleaved caspase-3. actin. Cytochrome C (10 M) Z-VAD-fmk. Procaspase-3. Cleaved caspase-3. actin. Z-VAD-fmk
A HeLa actin - + + - - + Cytochrome C (1 M) Z-VAD-fmk PMN - + + - - + actin Cytochrome C (1 M) Z-VAD-fmk Figure S1. (A) Pan-caspase inhibitor z-vad-fmk inhibits cytochrome c- mediated procaspase-3 cleavage.
More informationSupporting Information File S2
Pulli et al. Measuring Myeloperoxidase Activity in Biological Samples Page 1 of 6 Supporting Information File S2 Step-by-Step Protocol Reagents: Sucrose (Sigma, S3089) CaCl 2 (Sigma, C5770) Heparin Sodium
More informationab65336 Triglyceride Quantification Assay Kit (Colorimetric/ Fluorometric)
Version 10 Last updated 19 December 2017 ab65336 Triglyceride Quantification Assay Kit (Colorimetric/ Fluorometric) For the measurement of triglycerides in various samples. This product is for research
More informationSpontaneous Recanalization after Complete Occlusion of the Common Carotid Artery with Subsequent Embolic Ischemic Stroke
Original Contribution Spontaneous Recanalization after Complete Occlusion of the Common Carotid Artery with Subsequent Embolic Ischemic Stroke Abstract Introduction: Acute carotid artery occlusion carries
More informationCollagenase Assay Kit
Collagenase Assay Kit Catalog # 31 and 32 For Research Use Only - Not Human or Therapeutic Use INTRODUCTION The collagenases are members of the matrix metalloproteinase (MMP) family and degrade collagen
More informationSUPPLEMENTAL MATERIAL. Supplementary Methods
SUPPLEMENTAL MATERIAL Supplementary Methods Culture of cardiomyocytes, fibroblasts and cardiac microvascular endothelial cells The isolation and culturing of neonatal rat ventricular cardiomyocytes was
More informationTotal Phosphatidic Acid Assay Kit
Product Manual Total Phosphatidic Acid Assay Kit Catalog Number MET- 5019 100 assays FOR RESEARCH USE ONLY Not for use in diagnostic procedures Introduction Phosphatidic Acid (PA) is a critical precursor
More informationGUIDELINES FOR THE EARLY MANAGEMENT OF PATIENTS WITH ACUTE ISCHEMIC STROKE
2018 UPDATE QUICK SHEET 2018 American Heart Association GUIDELINES FOR THE EARLY MANAGEMENT OF PATIENTS WITH ACUTE ISCHEMIC STROKE A Summary for Healthcare Professionals from the American Heart Association/American
More informationHuman LDL ELISA Kit. Innovative Research, Inc.
Human LDL ELISA Kit Catalog No: IRKTAH2582 Lot No: SAMPLE INTRODUCTION Human low-density lipoprotein (LDL) transports cholesterol from the liver to tissues where it is incorporated into cell membranes.
More informationCerebrovascular Disease
Neuropathology lecture series Cerebrovascular Disease Physiology of cerebral blood flow Brain makes up only 2% of body weight Percentage of cardiac output: 15-20% Percentage of O 2 consumption (resting):
More informationIMAGING IN ACUTE ISCHEMIC STROKE
IMAGING IN ACUTE ISCHEMIC STROKE Timo Krings MD, PhD, FRCP (C) Professor of Radiology & Surgery Braley Chair of Neuroradiology, Chief and Program Director of Diagnostic and Interventional Neuroradiology;
More information11/27/2017. Stroke Management in the Neurocritical Care Unit. Conflict of interest. Karel Fuentes MD Medical Director of Neurocritical Care
Stroke Management in the Neurocritical Care Unit Karel Fuentes MD Medical Director of Neurocritical Care Conflict of interest None Introduction Reperfusion therapy remains the mainstay in the treatment
More informationVEGFR2-Mediated Vascular Dilation as a Mechanism of VEGF-Induced Anemia and Bone Marrow Cell Mobilization
Cell Reports, Volume 9 Supplemental Information VEGFR2-Mediated Vascular Dilation as a Mechanism of VEGF-Induced Anemia and Bone Marrow Cell Mobilization Sharon Lim, Yin Zhang, Danfang Zhang, Fang Chen,
More informationGlutathione S-Transferase Assay Kit
Glutathione S-Transferase Assay Kit Catalog Number KA1316 96 assays Version: 05 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Background... 3 Principle of the Assay...
More informationBY MARILYN M. RYMER, MD
Lytics, Devices, and Advanced Imaging The evolving art and science of acute stroke intervention. BY MARILYN M. RYMER, MD In 1996, when the US Food and Drug Administration (FDA) approved the use of intravenous
More informationIMAGING IN ACUTE ISCHEMIC STROKE
IMAGING IN ACUTE ISCHEMIC STROKE Timo Krings MD, PhD, FRCP (C) Professor of Radiology & Surgery Braley Chair of Neuroradiology, Chief and Program Director of Diagnostic and Interventional Neuroradiology;
More informationASSAY OF SPHINGOMYELINASE ACTIVITY
ASSAY OF SPHINGOMYELINASE ACTIVITY Protocol for Protein Extraction Stock Solution 1. Leupeptin/hydrochloride (FW 463.0,
More informationTissue type Plasminogen Activator Human Chromogenic Activity Assay Kit
ab108905 Tissue type Plasminogen Activator Human Chromogenic Activity Assay Kit Instructions for Use For the quantitative measurement of Human Tissue type Plasminogen Activator activity in cell culture
More informationMammalian Membrane Protein Extraction Kit
Mammalian Membrane Protein Extraction Kit Catalog number: AR0155 Boster s Mammalian Membrane Protein Extraction Kit is a simple, rapid and reproducible method to prepare cellular protein fractions highly
More informationWork-flow: protein sample preparation Precipitation methods Removal of interfering substances Specific examples:
Dr. Sanjeeva Srivastava IIT Bombay Work-flow: protein sample preparation Precipitation methods Removal of interfering substances Specific examples: Sample preparation for serum proteome analysis Sample
More informationAcoustic Pulse Thrombolysis Treatment
Acoustic Pulse Thrombolysis Treatment BTGVascular.com SETTING THE STANDARD FOR VASCULAR THERAPIES Quickly & safely dissolve thrombus with the EKOS System. The Acoustic Pulse Difference Acoustic Pulse Thrombolysis
More informationFOCUS SubCell. For the Enrichment of Subcellular Fractions. (Cat. # ) think proteins! think G-Biosciences
169PR 01 G-Biosciences 1-800-628-7730 1-314-991-6034 technical@gbiosciences.com A Geno Technology, Inc. (USA) brand name FOCUS SubCell For the Enrichment of Subcellular Fractions (Cat. # 786 260) think
More informationDiagnostic and Therapeutic Consequences of Repeat Brain Imaging and Follow-up Vascular Imaging in Stroke Patients
AJNR Am J Neuroradiol 0:7, January 999 Diagnostic and Therapeutic Consequences of Repeat Brain Imaging and Follow-up Vascular Imaging in Stroke Patients Birgit Ertl-Wagner, Tobias Brandt, Christina Seifart,
More informationCEREBRO VASCULAR ACCIDENTS
CEREBRO VASCULAR S MICHAEL OPONG-KUSI, DO MBA MORTON CLINIC, TULSA, OK, USA 8/9/2012 1 Cerebrovascular Accident Third Leading cause of deaths (USA) 750,000 strokes in USA per year. 150,000 deaths in USA
More informationRole of the Endothelial Surface Coat
S7 Role of the Endothelial Surface Coat in Microvascular Permeability Christopher Charles Michel Division of Biomedical Sciences, Imperial College School of Medicine, London, UK The luminal glycocalyx
More informationTotal Histone H3 Acetylation Detection Fast Kit (Colorimetric)
Total Histone H3 Acetylation Detection Fast Kit (Colorimetric) Catalog Number KA1538 48 assays Version: 02 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Intended Use...
More informationSensoLyte Generic MMP Assay Kit *Colorimetric*
SensoLyte Generic MMP Assay Kit *Colorimetric* Revision#1.2 Catalog # Kit Size Last updated: May2017 AS-72095 100 Assays (96-well plate) Optimized Performance: This kit is optimized to detect MMP activity
More informationChromatin IP (Isw2) Fix soln: 11% formaldehyde, 0.1 M NaCl, 1 mm EDTA, 50 mm Hepes-KOH ph 7.6. Freshly prepared. Do not store in glass bottles.
Chromatin IP (Isw2) 7/01 Toshi last update: 06/15 Reagents Fix soln: 11% formaldehyde, 0.1 M NaCl, 1 mm EDTA, 50 mm Hepes-KOH ph 7.6. Freshly prepared. Do not store in glass bottles. 2.5 M glycine. TBS:
More informationUse of EKOS Catheter in the management of Venous Mr. Manoj Niverthi, Mr. Sarang Pujari, and Ms. Nupur Dandavate, The GTF Group
Use of EKOS Catheter in the management of Venous Thromboembolism @ Mr. Manoj Niverthi, Mr. Sarang Pujari, and Ms. Nupur Dandavate, The GTF Group Introduction Georgia Thrombosis Forum (GTF, www.gtfonline.net)
More informationTECHNICAL BULLETIN. Catalog Number RAB0447 Storage Temperature 20 C
Phospho-Stat3 (ptyr 705 ) and pan-stat3 ELISA Kit for detection of human, mouse, or rat phospho-stat3 (ptyr 705 ) and pan-stat3 in cell and tissue lysates Catalog Number RAB0447 Storage Temperature 20
More informationHuman Leptin ELISA Kit
Product Manual Human Leptin ELISA Kit Catalog Numbers MET-5057 MET-5057-5 96 assays 5 x 96 assays FOR RESEARCH USE ONLY Not for use in diagnostic procedures Introduction Leptin is a polypeptide hormone
More informationThe Role of Neuroimaging in Acute Stroke. Bradley Molyneaux, HMS IV
The Role of Neuroimaging in Acute Stroke Bradley Molyneaux, HMS IV Patient CR 62 yo F w/ 2 wk h/o altered mental status Presents to ED w/ confusion following a fall 1 day prior New onset left facial droop
More informationCombination Studies With Tissue Plasminogen Activator
Effects of the Spin Trap Agent Disodium- [(tert-butylimino)methyl]benzene-1,3-disulfonate N-Oxide (Generic NXY-059) on Intracerebral Hemorrhage in a Rabbit Large Clot Embolic Stroke Model Combination Studies
More informationOcclusive cerebrovascular disease. A Novel Chronic Cerebral Hypoperfusion Model with Cognitive Impairment and Low Mortality Rate in Rats
FPⅧ-1 A Novel Chronic Cerebral Hypoperfusion Model with Cognitive Impairment and Low Mortality Rate in Rats Ahmed Said Mansour 1, Kuniyasu Niizuma 2, Sherif Rashad 2, Hidenori Endo 2, Toshiki Endo 3, Kenichi
More informationTECHNICAL BULLETIN. Phospho-Akt (pser 473 ) ELISA Kit for detection of human, mouse, or rat phospho-akt (pser 473 ) in cell and tissue lysates
Phospho-Akt (pser 473 ) ELISA Kit for detection of human, mouse, or rat phospho-akt (pser 473 ) in cell and tissue lysates Catalog Number RAB0011 Storage Temperature 20 C TECHNICAL BULLETIN Product Description
More informationACUTE ISCHEMIC STROKE. Current Treatment Approaches for Acute Ischemic Stroke
ACUTE ISCHEMIC STROKE Current Treatment Approaches for Acute Ischemic Stroke EARLY MANAGEMENT OF ACUTE ISCHEMIC STROKE Rapid identification of a stroke Immediate EMS transport to nearest stroke center
More informationWHITE PAPER: A GUIDE TO UNDERSTANDING SUBARACHNOID HEMORRHAGE
WHITE PAPER: A GUIDE TO UNDERSTANDING SUBARACHNOID HEMORRHAGE Subarachnoid Hemorrhage is a serious, life-threatening type of hemorrhagic stroke caused by bleeding into the space surrounding the brain,
More informationAdvanced age is an important risk factor for stroke and a
Longitudinal Magnetic Resonance Imaging of Sildenafil Treatment of Embolic Stroke in Aged Rats Guangliang Ding, PhD; Quan Jiang, PhD; Lian Li, PhD; Li Zhang, MD; Zhenggang Zhang, PhD, MD; Mei Lu, PhD;
More informationStroke Update. Lacunar 19% Thromboembolic 6% SAH 13% ICH 13% Unknown 32% Hemorrhagic 26% Ischemic 71% Other 3% Cardioembolic 14%
Stroke Update Michel Torbey, MD, MPH, FAHA, FNCS Medical Director, Neurovascular Stroke Center Professor Department of Neurology and Neurosurgery The Ohio State University Wexner Medical Center Objectives
More informationMatrix Metalloproteinases in Neuroinflammation
GLIA 39:279 291 (2002) Matrix Metalloproteinases in Neuroinflammation GARY A. ROSENBERG* Departments of Neurology, Neurosciences, and Cell Biology and Physiology, University of New Mexico Health Sciences
More informationSupporting Information for:
Supporting Information for: Methylerythritol Cyclodiphosphate (MEcPP) in Deoxyxylulose Phosphate Pathway: Synthesis from an Epoxide and Mechanisms Youli Xiao, a Rodney L. Nyland II, b Caren L. Freel Meyers
More informationBrain Attack. Strategies in the Management of Acute Ischemic Stroke: Neuroscience Clerkship. Case Medical Center
Brain Attack Strategies in the Management of Acute Ischemic Stroke: Neuroscience Clerkship Stroke is a common and devastating disorder Third leading antecedent of death in American men, and second among
More informationInhibition of MMP-9 by a selective gelatinase inhibitor protects neurovasculature from embolic focal cerebral ischemia
Cui et al. Molecular Neurodegeneration 2012, 7:21 RESEARCH ARTICLE Open Access Inhibition of MMP-9 by a selective gelatinase inhibitor protects neurovasculature from embolic focal cerebral ischemia Jiankun
More informationWestern Blot Analysis of Rat Pituitar Recognized by Human Antipituitary. y Antigens A. antibodies
Endocrine Journal 1995, 42(1), 115-119 NOTE Western Blot Analysis of Rat Pituitar Recognized by Human Antipituitary y Antigens A ntibodies SHIGEKI YABE, MASAMI MURAKAMI*, KAYOKO MARUYAMA, HIDEKO MIWA,
More informationCanadian Best Practice Recommendations for Stroke Care. (Updated 2008) Section # 3 Section # 3 Hyperacute Stroke Management
Canadian Best Practice Recommendations for Stroke Care (Updated 2008) Section # 3 Section # 3 Hyperacute Stroke Management Reorganization of Recommendations 2008 2006 RECOMMENDATIONS: 2008 RECOMMENDATIONS:
More informationStroke, the most common medical emergency, is a
Report on Progress 2016 Advances in Our Knowledge of Stroke Mechanisms and Therapy Xuefang Ren, M.D., and James W. Simpkins Ph. D Department of Physiology and Pharmacology, Experimental Stroke Core, Center
More informationHemodynamic Disorders, Thrombosis, and Shock. Richard A. McPherson, M.D.
Hemodynamic Disorders, Thrombosis, and Shock Richard A. McPherson, M.D. Edema The accumulation of abnormal amounts of fluid in intercellular spaces of body cavities. Inflammation and release of mediators
More informationLecithin Cholesterol Acyltransferase (LCAT) ELISA Kit
Product Manual Lecithin Cholesterol Acyltransferase (LCAT) ELISA Kit Catalog Number STA-616 96 assays FOR RESEARCH USE ONLY Not for use in diagnostic procedures Introduction Cholesterol is a lipid sterol
More informationMammalian Tissue Protein Extraction Reagent
Mammalian Tissue Protein Extraction Reagent Catalog number: AR0101 Boster s Mammalian Tissue Protein Extraction Reagent is a ready-to-use Western blot related reagent solution used for efficient extraction
More informationB16-F10 (Mus musculus skin melanoma), NCI-H460 (human non-small cell lung cancer
Electronic Supplementary Material (ESI) for ChemComm. This journal is The Royal Society of Chemistry 2017 Experimental Methods Cell culture B16-F10 (Mus musculus skin melanoma), NCI-H460 (human non-small
More informationStroke is a leading cause of death and disability worldwide,
Combination Treatment With VELCADE and Low-Dose Tissue Plasminogen Activator Provides Potent Neuroprotection in Aged Rats After Embolic Focal Ischemia Li Zhang, MD; Zheng Gang Zhang, MD, PhD; Ben Buller,
More informationeffect on the upregulation of these cell surface markers. The mean peak fluorescence intensity
SUPPLEMENTARY FIGURE 1 Supplementary Figure 1 ASIC1 disruption or blockade does not effect in vitro and in vivo antigen-presenting cell activation. (a) Flow cytometric analysis of cell surface molecules
More informationMichael Horowitz, MD Pittsburgh, PA
Michael Horowitz, MD Pittsburgh, PA Introduction Cervical Artery Dissection occurs by a rupture within the arterial wall leading to an intra-mural Hematoma. A possible consequence is an acute occlusion
More informationIndex. C Capillary telangiectasia, intracerebral hemorrhage in, 295 Carbon monoxide, formation of, in intracerebral hemorrhage, edema due to,
Neurosurg Clin N Am 13 (2002) 395 399 Index Note: Page numbers of article titles are in boldface type. A Age factors, in intracerebral hemorrhage outcome, 344 Albumin, for intracerebral hemorrhage, 336
More informationMatrix metalloproteinases (MMPs) belong to a large
Profiles of Matrix Metalloproteinases, Their Inhibitors, and Laminin in Stroke Patients Influence of Different Therapies Solveig Horstmann, MD; Pamela Kalb, MD; James Koziol, PhD; Humphrey Gardner, MD;
More informationHEART AND SOUL STUDY OUTCOME EVENT - MORBIDITY REVIEW FORM
REVIEW DATE REVIEWER'S ID HEART AND SOUL STUDY OUTCOME EVENT - MORBIDITY REVIEW FORM : DISCHARGE DATE: RECORDS FROM: Hospitalization ER Please check all that may apply: Myocardial Infarction Pages 2, 3,
More informationLongitudinal anterior-to-posterior shift of collateral channels in patients with moyamoya disease: an implication for its hemorrhagic onset
CLINICAL ARTICLE Longitudinal anterior-to-posterior shift of collateral channels in patients with moyamoya disease: an implication for its hemorrhagic onset Shusuke Yamamoto, MD, Satoshi Hori, MD, PhD,
More informationSystemic Inflammation Alters the Kinetics of Cerebrovascular Tight Junction Disruption after Experimental Stroke in Mice
The Journal of Neuroscience, September 17, 2008 28(38):9451 9462 9451 Neurobiology of Disease Systemic Inflammation Alters the Kinetics of Cerebrovascular Tight Junction Disruption after Experimental Stroke
More information