Donate Help Contact The AHA Sign In Home
American Heart Association
Stroke
Search: search_blue_button Advanced Search
« Previous Article | Table of Contents | Next Article »
Stroke. 2005;36:326-331
Published online before print January 6, 2005, doi: 10.1161/01.STR.0000152949.31366.3d
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
36/2/326    most recent
01.STR.0000152949.31366.3dv1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow Request Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Belayev, L.
Right arrow Articles by Ginsberg, M. D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Belayev, L.
Right arrow Articles by Ginsberg, M. D.
Related Collections
Right arrow Animal models of human disease
Right arrow Other Treatment
Right arrow Acute Cerebral Hemorrhage
Right arrow Behavioral Changes and Stroke
Right arrow Neuroprotectors

(Stroke. 2005;36:326.)
© 2005 American Heart Association, Inc.

Original Contributions

Albumin Treatment Reduces Neurological Deficit and Protects Blood–Brain Barrier Integrity After Acute Intracortical Hematoma in the Rat

Ludmila Belayev, MD; Isabel Saul, BS; Raul Busto, BS; Kristine Danielyan, PhD; Alexey Vigdorchik, MD; Larissa Khoutorova, BS Myron D. Ginsberg, MD

From the Cerebral Vascular Disease Research Center, Department of Neurology, University of Miami School of Medicine, Miami, Fla.

Correspondence to Dr Ludmila Belayev, Department of Neurology (D4–5), University of Miami School of Medicine, PO Box 016960, Miami, FL 33101. E-mail lbelayev{at}stroke.med.miami.edu


*    Abstract
up arrowTop
 *Abstract
down arrowIntroduction
 down arrowMaterials and Methods
 down arrowResults
 down arrowDiscussion
 down arrowConclusions
 down arrowReferences
 
Background and Purpose— Acute intracerebral hemorrhage (ICH) is a common and severe form of stroke. To date, medical management of ICH has had scant impact on morbidity and mortality. Because albumin therapy is markedly neuroprotective in preclinical models of ischemic stroke, and because ischemic and hemorrhagic stroke share several common injury mechanisms, we hypothesized that albumin therapy might also benefit ICH.

Methods— Acute intracortical hematoma was produced in anesthetized, normothermic rats by the single stereotaxic injection of 50 µL of autologous, nonheparinized whole blood over 5 minutes. Separate animal groups were treated either with 25% human albumin, 1.25 g/kg, or with intravenous saline vehicle at 60 minutes after ICH. Neurobehavior was quantified sequentially over the next 2 to 7 days. Damage to the blood–brain barrier was assessed at 2 days after ICH by fluorometric measurement of Evans blue extravasation in dissected brain regions.

Results— High-grade neurological deficits were present in all rats at 50 minutes after ICH (score 10.3±0.2, mean±SEM [maximal score 12]). Albumin-treated rats showed improved neuroscores relative to saline-treated animals beginning within hours of treatment and persisting throughout the 7-day survival period. At 3 and 7 days, mean total neuroscores of the albumin group were 38% to 43% lower than in saline-treated animals. Perihematomal Evans blue discoloration was readily evident in saline-treated ICH rats but was reduced by albumin treatment. Hemispheric Evans blue content ipsilateral to the hematoma was reduced by 49% by albumin treatment (albumin 93.9±13.3 versus saline 184.7±33.7 mg/g, P<0.05). Hematoma volume and brain swelling were not affected by albumin treatment.

Conclusions— Prompt albumin therapy improves neurological function and blood–brain barrier integrity after acute intracortical hematoma. These observations have important potential clinical implications.


Key Words: acute therapy • albumin • basic science • blood–brain barrier • neuroprotection • hemorrhage, intracranial • neuroprotective agents • neuroprotectants


*    Introduction
up arrowTop
 up arrowAbstract
 *Introduction
down arrowMaterials and Methods
 down arrowResults
 down arrowDiscussion
 down arrowConclusions
 down arrowReferences
 
Intracerebral hemorrhage (ICH) constitutes {approx}10% to 15% of all strokes and is associated with high morbidity and mortality. Thirty-day case fatality rates of {approx}35% have been reported in hospital-based studies and up to 52% in community-based studies,1 rates that are 2- to 6-fold higher than in ischemic stroke. Among chronic ICH survivors, nearly half are dependent on outside help in their activities of daily living, and only 20% of patients become independent at 6 months.2 To date, the medical management of ICH has had only a minimal impact on the disease, and the preclinical development of protective strategies has lagged markedly behind ischemic stroke.

Human albumin therapy, in moderate-to-high doses, has proved markedly neuroprotective in experimental models of focal3–6 and global7 cerebral ischemia and in traumatic brain injury.8 In the present study, we have extended the therapeutic applications of albumin therapy to the setting of ICH. The rationale for this approach is 2-fold: (1) ICH and ischemic stroke share a number of common injury mechanisms, which include perilesional edema and blood–brain barrier (BBB) disruption,9 oxidative injury and inflammation,10,11 excitotoxicity,12 and apoptosis.13 (2) Albumin therapy is known to antagonize many of these mechanisms by directly protecting both parenchymal and vascular elements of the brain, diminishing brain edema, maintaining microvascular integrity, inhibiting endothelial cell apoptosis, and exerting antioxidant effects.3–5,14 These considerations provide a strong rationale for suspecting that albumin therapy might also be beneficial in acute intracerebral hematoma. In the present study, we tested this hypothesis in a model of acute intracortical hematoma that gives rise to a consistent neurobehavioral deficit and BBB dysfunction. We present evidence that albumin therapy exerts a substantial protective effect in this setting.


*    Materials and Methods
up arrowTop
 up arrowAbstract
 up arrowIntroduction
 *Materials and Methods
down arrowResults
 down arrowDiscussion
 down arrowConclusions
 down arrowReferences
 
Animal Preparation
Twenty-nine male Sprague-Dawley rats (weight 289 to 378 g; Charles River Laboratories, Wilmington, Mass) were fasted overnight but allowed free access to water. Animal protocols for these studies were approved by the University of Miami Animal Care and Use Committee. Anesthesia was induced with 3.5% halothane in a mixture of 70% nitrous oxide and 30% oxygen. Rats were orally intubated and mechanically ventilated on 2% halothane and 70% nitrous oxide. Blood pressure arterial blood gases and pH were monitored, and left temporalis muscle temperature and rectal temperature were monitored with probes and maintained at 37.0°C to 37.5°C as described previously.5,15 Rectal temperature and body weight were monitored periodically after ICH.

Production of Intracortical Hematoma
Acute intracortical hemorrhage was produced by the single-injection method as described by Xue et al.16 In brief, the rat was placed in a stereotaxic frame (David Kopf Instruments), and a specially fabricated 27-gauge stainless steel cannula with 30° bevel was introduced through a burr hole into the midcerebral cortex (3 mm lateral to midline, 0.2 mm anterior to the coronal suture, and 2.5 mm below the surface of the skull). Each rat received a 50-µL injection of autologous whole blood over a period of 5 minutes with a microinfusion pump. (Blood was taken from the femoral artery with a 250-µL syringe, which was filled from the back of the barrel; no heparin was used.) After infusion, the injection cannula was left in place for 3 minutes and then removed slowly. The burr hole was filled with bone wax, the scalp incision closed, and the animal placed in a cage with free access to food and water.

Neurobehavioral Evaluation
A standardized quantitative neurobehavioral battery was used to assess sensorimotor function at 50 minutes after ICH and at sequential intervals after albumin or saline treatment. In 16 animals (albumin-treated, n=8; saline-treated, n=8), these observations were made at 2 hours, 4 hours, 1 day, and 2 days after treatment. Another 13 rats (albumin treated, n=6; saline-treated, n=7) were monitored from 1 hour through 7 days after treatment. The neurobehavioral battery consisted of postural reflex17 and placing tests,18 as described previously. Neurological function was graded on a scale of 0 to 11 (normal score=0, maximal score=12).15 Tests were conducted by an observer blinded to the treatment group.

Albumin Treatment
At 60 minutes after production of ICH, 14 rats received human serum albumin (25% solution, Baxter Healthcare Corp) 1.25 g/kg IV. Another 15 rats received a comparable volume of 0.9% saline vehicle.

Evaluation of BBB Integrity
We investigated the integrity of the BBB in 16 rats by measuring the extravasation of Evans blue.19,20 Evans blue dye (2% in saline, 4 mL/kg) was injected intravenously at 48 hours after ICH. Two hours later, the chest was opened under halothane anesthesia, and the brain was perfused with normal saline through the left ventricle at 110 mm Hg pressure until colorless perfusion fluid was obtained from the right atrium. After decapitation, the brain surface was photographed. The brain was then sectioned into 3 coronal blocks (2 mm in thickness) that included bregma levels +1.2, –0.3, and –1.3 mm, and the surfaces of each block were photographed.

In 8 brains, coronal blocks were next divided into right and left hemispheres and dissected into 8 regions for measurement of regional Evans blue (Figure 1). The dissected samples were weighed and extracted in 50% trichloroacetic acid solution. After homogenization and centrifugation, the extract was diluted with ethanol (1:3), and its fluorescence was determined (excitation at 620 nm and emission at 680 nm) with a Perkin-Elmer LS-5B luminescence spectrometer. Calculations were based on external standards in the same solvent (100 to 500 ng/mL). The tissue content of Evans blue was quantified from a linear standard curve derived from known amounts of the dye and was expressed per gram of tissue. We have described these methods previously.20



View larger version (31K):
[in this window]
[in a new window]
  		Figure 1. Coronal brain diagrams showing locations of intracortical hematoma and of tissue regions sampled for Evans blue assay. Coronal levels with reference to bregma are noted.

Histopathology
Two separate groups of animals (saline [n=6] and albumin [n=7]) were allowed to survive for 7 days. Brains were then perfusion fixed, paraffin embedded, coronally sectioned, and stained with hematoxylin and eosin as described previously.5,15 Hematoma area was measured at 9 standardized coronal levels by a blinded observer, and hematoma volume was computed by numerical integration with image-analysis methods reported previously.5 Brain swelling was determined from the difference in ipsilateral versus contralateral hemispheric volumes.

Statistical Analysis
Repeated-measures ANOVA was used for intergroup comparisons of neurobehavioral data and tissue Evans blue content.


*    Results
up arrowTop
 up arrowAbstract
 up arrowIntroduction
 up arrowMaterials and Methods
 *Results
down arrowDiscussion
 down arrowConclusions
 down arrowReferences
 
Physiological Variables
Rectal and cranial (temporalis muscle) temperatures, arterial blood pressure and blood gases, and plasma glucose levels in the 29 animals of this study showed no significant differences between groups. After treatment, modest hemodilution was present in the albumin group (36±1.2 versus 42±0.9 for the saline group).

Neurobehavior
Severe neurological deficits contralateral to ICH were evident in all rats when tested at 50 minutes after ICH (Figure 2). The neuroscores of albumin-treated rats tended to improve beyond those of the saline-treated group within 1 to 2 hours of treatment, and this intergroup difference was highly significant by 4 hours (P<0.01; Figure 2). Although progressive neurological improvement occurred in both groups over the 2- to 7-day survival period, the degree of recovery in albumin-treated animals exceeded that of the saline group at each time point (treatmentxtime interaction, P<0.01 by repeated-measures ANOVA; Figure 2). At 3 and 7 days, mean total neuroscores of the albumin group were 38% to 43% lower than in saline-treated animals.



View larger version (20K):
[in this window]
[in a new window]
  		Figure 2. Total neuroscores in rats with left intracortical hematoma before treatment (Pre-Rx) and at sequential time points after albumin or saline administration. Repeated-measures ANOVA revealed a significant overall between-subjects effect of albumin treatment (P<0.05) and a highly significant timextreatment interaction (P<0.01). *Post hoc Bonferroni comparison versus corresponding saline group, P<0.05.

Evans Blue Extravasation
Representative brains from each group are shown in Figure 3. A round hemorrhagic lesion was evident on the dorsolateral brain surface of all animals. Coronal sections revealed a cylindrical or triangular hemorrhagic lesion that extended through the neocortex with its epicenter at the coronal level of midstriatum (Figure 3). Brains of the saline-treated ICH group showed a halo of distinct Evans blue discoloration surrounding the hematoma, evident both on the brain surface and on coronal sections. This discoloration was fainter and less extensive in the albumin group (Figure 3). Spectrofluorometric measurement of tissue Evans blue content revealed marked left-hemisphere elevations, ipsilateral to the hematoma, which were highest in the cortex at the coronal level of the lesion itself (Figure 3; region L1 in Figure 4) and in the adjacent slice (region L2 in Figure 4). The albumin-treated group exhibited a marked reduction of overall left-hemisphere Evans blue extravasation, amounting to 49%, compared with saline-treated controls (Figure 4; P<0.05).



View larger version (52K):
[in this window]
[in a new window]
  		Figure 3. Coronal sections of representative saline- and albumin-treated animals with left intracortical hematoma and 2-day survival, administered Evans blue intravenously 2 hours before they were killed. Evans blue extravasation is grossly visible in the vehicle-treated animal, and substantial inhibition of the dye extravasation is evident in albumin-treated brain.



View larger version (17K):
[in this window]
[in a new window]
  		Figure 4. Tissue Evans blue content in saline- and albumin-treated rats with left intracortical hematoma and 2-day survival, measured in selected regions ipsilateral (left) and contralateral (right) to the hematoma (see Figure 1 for regions). Values are mean±SEM. *Different from saline group, P<0.01, Bonferroni test.

Histopathology
Histological examination after 7-day survival revealed a localized hematoma in the left cerebral cortex. Three animals showed hemorrhage in the adjacent white matter. Total hematoma volume was identical in the saline- and albumin-treated groups (2.11±0.7 and 2.07±0.25 mm3, respectively). Brain swelling was not affected by albumin treatment (saline-treated animals 0.5% and albumin-treated animals 1.1%, P=0.5, NS).


*    Discussion
up arrowTop
 up arrowAbstract
 up arrowIntroduction
 up arrowMaterials and Methods
 up arrowResults
 *Discussion
down arrowConclusions
 down arrowReferences
 
We have shown in this study that the prompt administration of moderate-dose human albumin to rats with acute cortical intracerebral hematoma leads to substantially improved neurological function throughout a 7-day survival period and to markedly improved BBB integrity as evidenced by diminished Evans blue permeability.

Vascular damage during cerebral ischemia occurs early and in a progressive fashion and contributes to edema, hemorrhagic transformation, and worsened clinical outcome in stroke patients.21 Although the beneficial effect of vascular protection has been shown in studies of cerebral ischemia, relatively few studies have addressed this topic in ICH models. Increased BBB permeability occurs after ICH and contributes to brain edema formation. During the early hours after ICH, the BBB remains intact to large molecules; however, BBB permeability increases markedly 8 to 12 hours later and even more so by 48 hours.22,23 Because edema is nearly maximal by 24 hours after ICH,23 therapy directed at reducing edema formation must be instituted within the first day. We have shown in the present study that prompt albumin therapy instituted at 60 minutes after production of ICH improves BBB integrity.

In cerebral ischemia, albumin-neuroprotection is mediated via multiple mechanisms. These include its major antioxidant actions,24 binding of transition metals,25 improvement of tissue perfusion,6,26 reversal of microvascular blood-element aggregation and sludging in pathological states,14 reduction of brain edema,3,4 normalization of brain water homeostasis,4 provision of essential fatty acids to the injured brain,27 and its role in maintaining normal endothelial28 and astrocytic function.29 In subjects with acute ischemic stroke, albumin therapy is currently being evaluated in a National Institutes of Health–supported phase I clinical trial (NS 40406).

In addition, albumin therapy might exert other beneficial effects specific to ICH itself. The pathophysiology of ICH involves not only mass effect and compressive injury to tissue but also potential toxicity from blood degradation products, which leads to secondary neuronal and parenchymal damage. A central event in ICH is the massive release of heme from red cells into the extracellular space9 and the metabolism of heme by heme oxygenase (HO).9 Free heme is both very hydrophobic and strongly pro-oxidant; it readily penetrates cell membranes, where it increases the susceptibility to oxidant-mediated killing. Heme also acts as a catalyst for the oxidation of LDL, which generates products that are toxic to endothelium.30 The release of heme also provides catalytically active iron to neighboring tissues.31 Albumin, together with hemopexin, is a major heme-binding protein. Albumin in the extracellular space is capable of binding free heme that results from hemorrhage or that is released from dying cells.32 The peroxidase- and catalase-like activities of heme are 50% to 60% inhibited by albumin. Once heme is complexed with albumin, it exhibits much lower reactivity toward hydrogen peroxide and other peroxides than nonprotein heme. Thus, by complexing heme, albumin helps to prevent the toxic effects of extracellular heme.32

Biliverdin is the initial product of HO-mediated hemoglobin cleavage and is subsequently reduced to bilirubin by biliverdin reductase.33 This enzyme is stabilized by serum albumin. Although heme itself is a potentially toxic pro-oxidant, biliverdin and bilirubin are both potent antioxidants.34,35 In its natural conformation, bilirubin is folded to yield a surface that contains an array of hydrophobic residues and hence is insoluble in aqueous media.36 Albumin binds to bilirubin with high affinity (2 moles per mole of albumin). Once bound to albumin, bilirubin protects albumin-bound fatty acids from peroxyl radical–induced oxidation.37 These mechanisms may contribute to the albumin-induced neuroprotection observed in the present study.

In the present study, there were no differences among treatment groups with respect to hematoma size or brain swelling, but neurobehavioral outcome was significantly improved by albumin therapy. ICH differs from ischemic stroke in that the volume of tissue into which bleeding occurs may be irretrievably lost, even with early intervention. The results described here are consistent with other studies in suggesting that behavioral measures may be more responsive to therapy than histopathology. For example, treatment with the free radical inhibitors alpha-phenyl-N-tert-butyl nitrone38 and NXY-05939 significantly improved behavioral function in rats after ICH but did not affect hematoma size or edema.

The model of acute intracortical hematoma used in the present study gives rise to a severe neurobehavioral deficit together with consistent BBB dysfunction. The model is relevant to the clinical condition of lobar hematoma.40 Thrombin, a serine protease essential to the coagulation cascade, plays a major role in inducing early edema and BBB disruption, and the use of heparinized blood to induce ICH may impede this injury mechanism.41 Therefore, in the present study, we used nonheparinized blood. Of relevance in this respect, in stroke patients treated with tissue plasminogen activator who develop thrombolysis-related ICH, the degree of perihematomal edema is much less than in patients with spontaneous ICH.42 This observation implicates intrahematomal blood clotting as a major pathogenetic factor in hyperacute perihematomal edema.42


*    Conclusions
up arrowTop
 up arrowAbstract
 up arrowIntroduction
 up arrowMaterials and Methods
 up arrowResults
 up arrowDiscussion
 *Conclusions
down arrowReferences
 
We have shown that prompt therapy with moderate-dose human albumin reduces neurological deficits and improves BBB integrity in the setting of acute intracortical hematoma in the rat. These findings, once confirmed and extended, have important potential clinical implications. One intriguing implication of the fact that human albumin therapy is neuroprotective in both acute ischemic stroke and acute ICH is that this therapy, if validated in controlled clinical trials, could be instituted in acute stroke patients without the need for a prior CT scan to distinguish ischemic from hemorrhagic stroke.


*    Acknowledgments
 
This study was supported by National Institutes of Health program-project grant NS 05820. The authors thank Guillermo Fernandez, BS, for his technical assistance.

Received October 5, 2004; revision received October 22, 2004; accepted October 27, 2004.


*    References
up arrowTop
 up arrowAbstract
 up arrowIntroduction
 up arrowMaterials and Methods
 up arrowResults
 up arrowDiscussion
 up arrowConclusions
 *References
 

  1. Norrving, B. Cerebral hemorrhage. In: Ginsberg, MD, Bogousslavsky, J, eds. Cerebrovascular Disease: Pathophysiology, Diagnosis, and Management. Malden, Mass: Blackwell Science; 1998.
  2. Fogelholm R, Nuutila M, Vuorela AL. Primary intracerebral haemorrhage in the Jyvaskyla region, central Finland, 1985–89: incidence, case fatality rate, and functional outcome. J Neurol Neurosurg Psychiatry. 1992; 55: 546–552.[Abstract]
  3. Belayev L, Busto R, Zhao W, Clemens JA, Ginsberg MD. Effect of delayed albumin hemodilution on infarction volume and brain edema after transient middle cerebral artery occlusion in rats. J Neurosurg. 1997; 87: 595–601.[Medline] [Order article via Infotrieve]
  4. Belayev L, Zhao W, Pattany PM, Weaver RG, Huh PW, Lin B, Busto R, Ginsberg MD. Diffusion-weighted magnetic resonance imaging confirms marked neuroprotective efficacy of albumin therapy in focal cerebral ischemia. Stroke. 1998; 29: 2587–2599.[Abstract/Free Full Text]
  5. Belayev L, Liu Y, Zhao W, Busto R, Ginsberg MD. Human albumin therapy of acute ischemic stroke: marked neuroprotective efficacy at moderate doses and with a broad therapeutic window. Stroke. 2001; 32: 553–560.[Abstract/Free Full Text]
  6. Liu Y, Belayev L, Zhao W, Busto R, Belayev A, Ginsberg MD. Neuroprotective effect of treatment with human albumin in permanent focal cerebral ischemia: histopathology and cortical perfusion studies. Eur J Pharmacol. 2001; 428: 193–201.[CrossRef][Medline] [Order article via Infotrieve]
  7. Belayev L, Saul I, Huh PW, Finotti N, Zhao W, Busto R, Ginsberg MD. Neuroprotective effect of high-dose albumin therapy against global ischemic brain injury in rats. Brain Res. 1999; 845: 107–111.[CrossRef][Medline] [Order article via Infotrieve]
  8. Belayev L, Alonso OF, Huh PW, Zhao W, Busto R, Ginsberg MD. Posttreatment with high-dose albumin reduces histopathological damage and improves neurological deficit following fluid percussion brain injury in rats. J Neurotrauma. 1999; 16: 445–453.[Medline] [Order article via Infotrieve]
  9. Wagner KR, Sharp FR, Ardizzone TD, Lu A, Clark JF. Heme and iron metabolism: role in cerebral hemorrhage. J Cereb Blood Flow Metab. 2003; 23: 629–652.[CrossRef][Medline] [Order article via Infotrieve]
  10. Wagner KR, Packard BA, Hall CL, Smulian AG, Linke MJ, De Court, Packard LM, Hall NC. Protein oxidation and heme oxygenase-1 induction in porcine white matter following intracerebral infusions of whole blood or plasma. Dev Neurosci. 2002; 24: 154–160.[CrossRef][Medline] [Order article via Infotrieve]
  11. Xue M, Del Bigio MR. Acute tissue damage after injections of thrombin and plasmin into rat striatum. Stroke. 2001; 32: 2164–2169.[Abstract/Free Full Text]
  12. Qureshi AI, Ali Z, Suri MF, Shuaib A, Baker G, Todd K, Guterman LR, Hopkins LN. Extracellular glutamate and other amino acids in experimental intracerebral hemorrhage: an in vivo microdialysis study. Crit Care Med. 2003; 31: 1482–1489.[CrossRef][Medline] [Order article via Infotrieve]
  13. Matsushita K, Meng W, Wang X, Asahi M, Asahi K, Moskowitz MA, Lo EH. Evidence for apoptosis after intercerebral hemorrhage in rat striatum. J Cereb Blood Flow Metab. 2000; 20: 396–404.[CrossRef][Medline] [Order article via Infotrieve]
  14. Belayev L, Pinard E, Nallet H, Seylaz J, Liu Y, Riyamongkol P, Zhao W, Busto R, Ginsberg MD. Albumin therapy of transient focal cerebral ischemia: in vivo analysis of dynamic microvascular responses. Stroke. 2002; 33: 1077–1084.[Abstract/Free Full Text]
  15. Belayev L, Alonso OF, Busto R, Zhao W, Ginsberg MD. Middle cerebral artery occlusion in the rat by intraluminal suture: neurological and pathological evaluation of an improved model. Stroke. 1996; 27: 1616–1622.[Abstract/Free Full Text]
  16. Xue M, Del Bigio MR. Intracortical hemorrhage injury in rats: relationship between blood fractions and brain cell death. Stroke. 2000; 31: 1721–1727.[Abstract/Free Full Text]
  17. 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: 472–476.[Abstract/Free Full Text]
  18. De Ryck M, Van Reempts J, Borgers M, Wauquier A, Janssen PJ. Photochemical stroke model: flunarizine prevents sensorimotor deficits after neocortical infarcts in rats. Stroke. 1989; 20: 1383–1390.[Abstract/Free Full Text]
  19. Uyama O, Okamura N, Yanase M, Narita M, Kawabata K, Sugita M. Quantitative evaluation of vascular permeability in the gerbil brain after transient ischemia using Evans blue fluorescence. J Cereb Blood Flow Metab. 1988; 8: 282–284.[Medline] [Order article via Infotrieve]
  20. Belayev L, Busto R, Zhao W, Ginsberg MD. Quantitative evaluation of blood-brain barrier permeability following middle cerebral artery occlusion in rats. Brain Res. 1996; 739: 88–96.[CrossRef][Medline] [Order article via Infotrieve]
  21. Fagan SC, Hess DC, Hohnadel EJ, Pollock DM, Ergul A. Targets for vascular protection after acute ischemic stroke. Stroke. 2004; 35: 2220–2225.[Abstract/Free Full Text]
  22. Wagner KR, Xi G, Hua Y, Kleinholz M, Courten-Myers GM, Myers RE, Broderick JP, Brott TG. Lobar intracerebral hemorrhage model in pigs: rapid edema development in perihematomal white matter. Stroke. 1996; 27: 490–497.[Abstract/Free Full Text]
  23. Yang GY, Betz AL, Chenevert TL, Brunberg JA, Hoff JT. Experimental intracerebral hemorrhage: relationship between brain edema, blood flow, and blood-brain barrier permeability in rats. J Neurosurg. 1994; 81: 93–102.[Medline] [Order article via Infotrieve]
  24. Halliwell B, Gutteridge JM. The antioxidants of human extracellular fluids. Arch Biochem Biophys. 1990; 280: 1–8.[CrossRef][Medline] [Order article via Infotrieve]
  25. Emerson TE. Unique features of albumin: a brief review. Crit Care Med. 1989; 17: 690–694.[Medline] [Order article via Infotrieve]
  26. Huh PW, Belayev L, Zhao W, Busto R, Saul I, Ginsberg MD. The effect of high-dose albumin therapy on local cerebral perfusion after transient focal cerebral ischemia in rats. Brain Res. 1998; 804: 105–113.[CrossRef][Medline] [Order article via Infotrieve]
  27. Rodriguez de Turco EB, Belayev L, Liu Y, Busto R, Parkins N, Bazan NG, Ginsberg MD. Systemic fatty acid responses to transient focal cerebral ischemia: influence of neuroprotectant therapy with human albumin. J Neurochem. 2002; 83: 515–524.[CrossRef][Medline] [Order article via Infotrieve]
  28. Keaney JFJ, Simon DI, Stamler JS, Jaraki O, Scharfstein J, Vita JA, Loscalzo J. NO forms an adduct with serum albumin that has endothelium-derived relaxing factor-like properties. J Clin Invest. 1993; 91: 1582–1589.[Medline] [Order article via Infotrieve]
  29. Tabernero A, Medina A, Sanchez-Abarca LI, Lavado E, Medina JM. The effect of albumin on astrocyte energy metabolism is not brought about through the control of cytosolic Ca2+ concentrations but by free-fatty acid sequestration. Glia. 1999; 25: 1–9.[CrossRef][Medline] [Order article via Infotrieve]
  30. Jeney V, Balla J, Yachie A, Varga Z, Vercellotti GM, Eaton JW, Balla G. Pro-oxidant and cytotoxic effects of circulating heme. Blood. 2002; 100: 879–887.[Abstract/Free Full Text]
  31. Balla J, Jacob HS, Balla G, Nath K, Eaton JW, Vercellotti GM. Endothelial-cell heme uptake from heme proteins: induction of sensitization and desensitization to oxidant damage. Proc Natl Acad Sci U S A. 1993; 90: 9285–9289.[Abstract/Free Full Text]
  32. Grinberg LN, O’Brien PJ, Hrkal Z. The effects of heme-binding proteins on the peroxidative and catalatic activities of hemin. Free Radic Biol Med. 1999; 27: 214–219.[CrossRef][Medline] [Order article via Infotrieve]
  33. Kutty RK, Maines MD. Purification and characterization of biliverdin reductase from rat liver. J Biol Chem. 1981; 256: 3956–3962.[Abstract/Free Full Text]
  34. Dore S, Takahashi M, Ferris CD, Zakhary R, Hester LD, Guastella D, Snyder SH. Bilirubin, formed by activation of heme oxygenase-2, protects neurons against oxidative stress injury. Proc Natl Acad Sci U S A. 1999; 96: 2445–2450.[Abstract/Free Full Text]
  35. Baranano DE, Rao M, Ferris CD, Snyder SH. Biliverdin reductase: a major physiologic cytoprotectant. Proc Natl Acad Sci U S A. 2002; 99: 16093–16098.[Abstract/Free Full Text]
  36. Smith LH Jr, Their SO. Pathophysiology. The Biological Principles of Disease. Philadelphia, Pa: WB Saunders; 1985.
  37. Stocker R, Glazer AN, Ames BN. Antioxidant activity of albumin-bound bilirubin. Proc Natl Acad Sci U S A. 1987; 84: 5918–5922.[Abstract/Free Full Text]
  38. Peeling J, Yan HJ, Chen SG, Campbell M, Del Bigio MR. Protective effects of free radical inhibitors in intracerebral hemorrhage in rat. Brain Res. 1998; 795: 63–70.[CrossRef][Medline] [Order article via Infotrieve]
  39. Peeling J, Del Bigio MR, Corbett D, Green AR, Jackson DM. Efficacy of disodium 4-[(tert-butylimino)methyl]benzene-1,3-disulfonate N-oxide (NXY-059), a free radical trapping agent, in a rat model of hemorrhagic stroke. Neuropharmacology. 2001; 40: 433–439.[CrossRef][Medline] [Order article via Infotrieve]
  40. Labovitz DL, Sacco RL. Intracerebral hemorrhage: update. Curr Opin Neurol. 2001; 14: 103–108.[CrossRef][Medline] [Order article via Infotrieve]
  41. Xi G, Wagner KR, Keep RF, Hua Y, Courten-Myers GM, Broderick JP, Brott TG, Hoff JT, Muizelaar JP. Role of blood clot formation on early edema development after experimental intracerebral hemorrhage. Stroke. 1998; 29: 2580–2586.[Abstract/Free Full Text]
  42. Gebel JM, Brott TG, Sila CA, Tomsick TA, Jauch E, Salisbury S, Khoury J, Miller R, Pancioli A, Duldner JE, Topol EJ, Broderick JP. Decreased perihematomal edema in thrombolysis-related intracerebral hemorrhage compared with spontaneous intracerebral hemorrhage. Stroke. 2000; 31: 596–600.[Abstract/Free Full Text]



This article has been cited by other articles:


Home page
 Cancer Res.Home page
M. Veiseh, P. Gabikian, S-B. Bahrami, O. Veiseh, M. Zhang, R. C. Hackman, A. C. Ravanpay, M. R. Stroud, Y. Kusuma, S. J. Hansen, et al.
Tumor Paint: A Chlorotoxin:Cy5.5 Bioconjugate for Intraoperative Visualization of Cancer Foci
Cancer Res., July 15, 2007; 67(14): 6882 - 6888.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
36/2/326    most recent
01.STR.0000152949.31366.3dv1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow Request Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Belayev, L.
Right arrow Articles by Ginsberg, M. D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Belayev, L.
Right arrow Articles by Ginsberg, M. D.
Related Collections
Right arrow Animal models of human disease
Right arrow Other Treatment
Right arrow Acute Cerebral Hemorrhage
Right arrow Behavioral Changes and Stroke
Right arrow Neuroprotectors

Stroke Home | Subscriptions | Archives | Feedback | Authors | Help | AHA Journals Home | Search
Copyright © 2005 American Heart Association, Inc. All rights reserved. Unauthorized use prohibited.