Primate Stroke Model Research Articles

NXY-059, the free radical trapping nitrone-based agent, reduces hemiparesis and reduces grey and white matter damage after focal cerebral ischemia in monkeys

60 There is little published evidence for protection of white matter with neuroprotective drugs in animal models of stroke, yet white matter protection may be important in achieving clinical efficacy. We have examined the effects of NXY-059, a nitrone-based free radical trapping agent, on long-term functional disability in a primate model of stroke. We also examined histopathological effects, including analyses of grey and white matter damage. Five minutes after unilateral permanent middle cerebral artery occlusion, marmosets received a 1 ml i.v. infusion of saline (n=5) or NXY-059 (28 mg/kg) (n=6) and osmotic minipumps (model 2001D) were implanted s.c. to provide continuous drug or saline infusion for 48 h. Drug-filled pumps released NXY-059 at a rate of approximately 16 mg/kg/h. The plasma unbound drug concentration at 24 h was 76.3 ± 5.7 μM, a level well tolerated in acute stroke patients. The monkeys had been trained and tested on a variety of behavioral tasks before surgery. NXY-059-treated monkeys were significantly better at reaching with their contralesional arm than were saline-treated monkeys when re-tested 3 (p<0.01) and 10 weeks (p<0.01) after surgery. NXY-059-treatment also significantly reduced spatial neglect measured 3 weeks after surgery (p<0.01) compared with the saline group. After behavioral testing was complete, histopathological analysis showed NXY-059-treated monkeys had significantly smaller infarcts than saline-treated monkeys (F (1, 10)=5.21, p<0.05). NXY-059 reduced overall infarct size by 51%, damage to the cortex was reduced by 54%, white matter by 52%, caudate by 49%, and putamen by 33% compared with saline-treated monkeys. In conclusion, NXY-059 substantially lessened the functional disability in these monkeys. This drug protects not only cortical tissue, but also white matter and subcortical structures against ischemic damage. Together these findings bode well for advancing this drug to further clinical trials for its use in acute stroke.

NXY-059, the free radical trapping nitrone-based agent, reduces hemiparesis and reduces grey and white matter damage after focal cerebral ischemia in monkeys

60 There is little published evidence for protection of white matter with neuroprotective drugs in animal models of stroke, yet white matter protection may be important in achieving clinical efficacy. We have examined the effects of NXY-059, a nitrone-based free radical trapping agent, on long-term functional disability in a primate model of stroke. We also examined histopathological effects, including analyses of grey and white matter damage. Five minutes after unilateral permanent middle cerebral artery occlusion, marmosets received a 1 ml i.v. infusion of saline (n=5) or NXY-059 (28 mg/kg) (n=6) and osmotic minipumps (model 2001D) were implanted s.c. to provide continuous drug or saline infusion for 48 h. Drug-filled pumps released NXY-059 at a rate of approximately 16 mg/kg/h. The plasma unbound drug concentration at 24 h was 76.3 ± 5.7 μM, a level well tolerated in acute stroke patients. The monkeys had been trained and tested on a variety of behavioral tasks before surgery. NXY-059-treated monkeys were significantly better at reaching with their contralesional arm than were saline-treated monkeys when re-tested 3 (p

NXY-059, a Free Radical–Trapping Agent, Substantially Lessens the Functional Disability Resulting From Cerebral Ischemia in a Primate Species

NXY-059 is a novel nitrone with free radical-trapping properties that has a considerable neuroprotective effect in rats. We have now examined the efficacy of this drug at reducing long-term functional disability in a primate model of stroke. Twelve monkeys were trained and tested on a variety of behavioral tasks used to dissociate and quantify motor and spatial deficits. Five minutes after permanent occlusion of the right middle cerebral artery, monkeys received a 1-mL intravenous infusion of either saline or NXY-059 (28 mg x kg(-1)), and osmotic minipumps, model 2001D, were implanted subcutaneously to provide continuous drug or saline infusion for 48 hours. Drug-filled pumps released NXY-059 at 16 mg x kg(-1) x h(-1). The monkeys were retested 3 and 10 weeks after surgery to assess functional disability. Surgery, behavioral testing, and histology were all done blinded to treatment condition. NXY-059-treated monkeys were significantly better at reaching with their hemiparetic arm than were saline-treated monkeys when retested 3 weeks (P:<0.01) and 10 weeks (P:<0.01) after surgery. Drug treatment also significantly lessened the degree of spatial perceptual neglect (P:<0.01), a debilitating though ameliorating consequence of this infarct. NXY-059 treatment reduced the overall amount of brain damage by >50% of saline-treatment values, with similar levels of protection afforded to both white and gray matter. This novel drug has a substantial protective effect, lessening the disability caused by an experimentally induced stroke in a primate species. These findings provide considerable encouragement for the clinical development of NXY-059.

AR-R15896AR, a low affinity, use-dependent, NMDA antagonist, is protective in a primate model of stroke

The low affinity, use-dependent, N-methyl- D-aspartate (NMDA) antagonist, AR-R15896AR, is neuroprotective against transient focal cerebral ischemia in rats. We have examined the effect of AR-R15896AR, administered at a plasma level that is tolerated in acute stroke patients, on both functional and histopathologic measures in marmoset monkeys with permanent middle cerebral artery (MCA) occlusion. The M1 segment of the right MCA was permanently occluded (pMCAO) by bipolar coagulation in 11 marmosets. Five minutes later, the monkeys received either saline or AR-R15896 (4.5 mg/kg) intravenously, infused over 30 seconds. Also, osmotic minipumps were implanted subcutaneously to provide continuous drug or saline infusion for 48 hours. Drug-filled pumps released AR-R15896 at a rate of 1.1 mg/kg/h. The monkeys had been trained and tested preoperatively on a number of behavioral tasks and were retested 3 and 10 weeks after surgery. Three weeks after surgery, all the monkeys had a severe motor deficit, in that they were impaired at reaching with their contralesional arms, and perceptual neglect of contralesional space. AR-R15896AR-treated monkeys, however, had significantly less neglect than saline-treated monkeys, although the AR-R15896AR treatment had no effect on the motor deficit. By 10 weeks, the neglect had recovered, but not the motor deficit, and there were no differences between the 2 groups. Histopathologic analysis showed a reduction in the size of the infarction at several stereotaxic levels of the AR-R15896AR-treated monkeys. This study has shown that AR-R15896AR protected a select region of the brain against permanent focal cerebral ischemia in a primate species and attenuated the ensuing spatial neglect.

Clomethiazole protects against hemineglect in a primate model of stroke

Permanent occlusion of the M1 segment of the middle cerebral artery (pMCAO) in the marmoset, a New World species of monkey, produces unilateral functional deficits, including motor neglect with the contralesional arm and contralesional spatial hemineglect. In this study we examined whether clomethiazole, a drug which modulates the γ-aminobutyric acid A receptor, reduced the severity of the hemineglect and other deficits in this primate model of stroke. Nine monkeys received pMCAO; 1 h later four of the nine were administered clomethiazole by intraperitoneal injection and subcutaneous implantation of osmotic mini-pumps, which released clomethiazole for 48 h. The monkeys had been trained and tested on a number of behavioral tasks prior to surgery and were re-tested 3 and 10 weeks later. Three weeks after pMCAO, monkeys treated with clomethiazole had a significantly reduced degree of spatial neglect compared to untreated controls. Clomethiazole was not effective against the severe contralesional motor impairment in the current study, although it ameliorated a somewhat less severe motor deficit in a previous study in which the more distal, M2 segment of the middle cerebral artery had been occluded. Postmortem analysis of the brains showed that clomethiazole treatment had significantly reduced the area of damage in part of the parietal cortex. These data suggest that clomethiazole may reduce the neglect that can be a debilitating consequence of right-sided stroke in man.

Comprehensive MR imaging protocol for stroke management: tissue sodium concentration as a measure of tissue viability in nonhuman primate studies and in clinical studies.

To investigate sodium magnetic resonance (MR) imaging for monitoring tissue viability in stroke. A comprehensive MR imaging protocol used to measure apparent diffusion coefficient and perfusion parameters was extended to include sodium imaging. Tissue sodium concentration was estimated by using a two-compartment model. This protocol lasted less than 45 minutes. These parameters were followed over the first 6 hours in a nonhuman primate model (n = 2) of acute embolic stroke without or with thrombolytic therapy. This protocol was used in patients in whom acute (< 24 hours, n = 11) or nonacute (> or = 24 hours, n = 31) stroke was ultimately confirmed. The animal model showed abnormal diffusion and perfusion parameters in the lesion immediately after embolization, and these remained abnormal for over 6 hours. Tissue sodium concentration increased with time (5.7 mmol/L/h) unless halted with thrombolytic therapy. Regions with sodium concentrations over 70 mmol/L were histochemically verified as being infarcted. In patients in whom stroke older than 6 hours was clinically confirmed, sodium concentrations over 70 mmol/L were found in the appropriate brain regions. Tissue sodium concentration provides a sensitive measure of tissue viability that is complementary to the diagnostic role of diffusion and perfusion imaging for ischemic insult.

Effect of Fluosol-DA and hetastarch on local cerebral blood flow, cortical O2 availability and computerized EEG data during cerebral ischaemia*

This study was designed to assess the effects of volume expansion with Fluosol-DA and hetastarch on local cerebral blood flow (CBF), cortical O2 availability (O2a) and computerized EEG power data during cerebral ischaemia in a primate stroke model. Focal cerebral ischaemia was produced in 20 anaesthetized Macaca monkeys by right unilateral middle cerebral artery (MCA) occlusion by the transorbital technique. Following occlusion of the right MCA, monkeys were randomized into one of the following treatment groups: Group A--Fluosol-DA 15 ml kg-1 i.v. + 40% O2; Group B--Fluosol-DA 15 ml kg-1 i.v. + 100% O2; Group C--hetastarch 30 ml kg-1 i.v. + 40% O2; Group D--hetastarch 30 ml kg-1 i.v. + 100% O2. In Group A, local CBF increased and EEG power data improved while O2a showed no change in the right MCA territory. In Group B, local CBF increased from 84% (P less than 0.001), O2a improved significantly, and EEG power data showed significant improvement in the right hemisphere. Cardiac output did not change during the Fluosol-DA infusions. In Groups C and D, local CBF increased significantly, O2a did not change, and EEG power data improved significantly in the right hemisphere. Cardiac output also increased significantly during the hetastarch infusions. These results show that Fluosol-DA and hetastarch improve local CBF and EEG power data in ischaemic brain. Only the Fluosol-DA + 100% O2 increased the O2a in the ischaemic brain. We conclude that the benefits with Fluosol-DA are due to its ability to increase O2 delivery to ischaemic brain.

Temporal profile of tissue levels of dopamine and its metabolites, HVA and DOPAC following focal cerebral ischaemia in anaesthetized primates.

Occlusion of the middle cerebral artery produced ischaemia and consequent changes in basal ganglia in the primate model of stroke within 0.5 h. Dopamine content was decreased in the right basal ganglia from 0.5 h up to 12 h after occlusion. Homovanillic acid content of the right basal ganglia increased initially at 0.5 h but, was decreased at 12 h. The DOPAC content was increased at 2 h after occlusion in the right basal ganglia, but was decreased significantly at 12 h. In the substantia nigra, there was a significant reduction in the DA content in the right side at 12 h. Minimal changes were observed in the DOPAC and HVA content in the right substantia nigra at 2 h and 4 h, respectively. At other time periods there was no significant change in the content of HVA and DOPAC in substantia nigra.

Effect of Naloxone on Experimental Stroke in Awake Monkeys

Naloxone has recently been reported to be of benefit in the treatment of central nervous system ischemia. To study the effect of naloxone in an experimental stroke model, we subjected 40 awake monkeys to middle cerebral artery (MCA) occlusion 2 weeks after the placement of a MCA ligature by a transorbital technique. Cerebral blood flow (CBF) was monitored with stereotactically placed H2 electrodes, and the neurological course was serially recorded. Infarct size was determined 2 weeks after MCA occlusion. Twenty animals served as control and received no naloxone; 10 of these underwent permanent occlusion, and 10 underwent 4-hour MCA occlusion. In 25 treatment animals, naloxone was administered in three different intravenous dosages: (a) naloxone, 2-mg/kg bolus 20 minutes postocclusion and 2 mg/kg/hour X 4 hours, in 10 animals with permanent MCA occlusion; (b) naloxone, 10-mg/kg bolus 20 minutes postocclusion and 10 mg/kg/hour X 4 hours, in 10 animals with 4-hour MCA occlusion; and (c) naloxone, 20-mg/kg bolus, in 5 animals with various neurological deficits. MCA occlusion typically produced a moderate deficit: hemiparesis, hemianopsia, and facial paresis. In most instances, naloxone in the 2- and 10-mg/kg dose regimens produced little or no change in the neurological function. CBF decreased after MCA occlusion and was unaffected by naloxone in most cases. Infarct size was not significantly different between the control and treated groups. However, the 20-mg/kg dose consistently produced a nonfunctional, transient increase in total body motor tone in normal and hemiparetic animals. Naloxone did not significantly improve useful neurological function, CBF, or infarct size in an experimental primate stroke model.

Beneficial effects of induced hypertension on experimental stroke in awake monkeys.

The authors performed a controlled study of induced hypertension therapy for treatment of experimental stroke in unanesthetized monkeys. Ten control and 10 treated animals were subjected to a 4-hour occlusion of the middle cerebral artery (MCA) by an implanted tourniquet. Neurological status and local cerebral blood flow (CBF) were monitored serially. Local CBF was determined by hydrogen clearance in and around the elevated 20% to 40% by intravenous infusion of phenylephrine hydrochloride. Neuropathological evaluation was performed after about 2 weeks. A 4-hour occlusion of the MCA in control animals caused moderate stable neurological deficits, moderate stable decreases in local CBF, and medium-sized infarcts. With induced hypertension, five of 10 treated animals showed neurological improvement, and eight exhibited increased CBF in the ischemic zone. Average infarct size tended to be smaller in the treated group, although the difference did not reach statistical significance. Hemorrhagic infarcts were not observed. In four animals, phenylephrine caused cardiac dysrhythmias and hypotension which were reversed by appropriate measures. In this unanesthetized primate model of moderate experimental stroke, induced hypertension had beneficial effects on neurological status, local CBF, and infarct size without causing hemorrhagic infarction. Induced hypertension may be beneficial for some clinical cases of focal cerebral ischemia.

Epidemiology, emergency and acute care: Advances in physiopathology and treatment

is Assistant Professor of Neurological Surgery, University of Pittsburgh, He received his basic neurosurgery training at Case Western Reserve Hospital and completed a year Fellowship in Microsurgery at the University of Pittsburgh. Dr. Yonas's research areas mainly are centered on problems of cerebral ischemia; for that study he has developed a new primate stroke model. His special interests in problems of spasticity developed during his fellowship at the University of Pittsburgh, and continues to the present.

Alterations in cortical oxygen tension during the development of ischemic cerebral edema in primates (Macaca mulatta).

With a closed head primate stroke model, acute cerebral ischemia limited to the middle cerebral artery (MCA) territory was produced by macrosphere embolization of the internal carotid artery bifurcation. Measurements of the oxygen tension (PO2) at the cerebral cortical surface were obtained by continuous on-line mass spectrometry. Percentage of dry weight and tissue sodium, potassium, and chloride concentrations from ischemic and nonischemic hemispheres were determined at various times. With this preparation, we registered the precise onset of cortical surface PO2 depletion, which showed an exponential downward trend (fast component from 0 to 5 minutes, t 1/2 = 0.8 minute, rate of change = 89% per minute; slow component from 5 to 240 minutes, t 1/2 = 285 minutes, rate of change = 0.3% per minute). After the onset of cerebral ischemia, there was an immediate fall of the cortical surface PO2 with reductions of more than 45% at 5 minutes before definite hemiparesis and electroencephalographic abnormalities were recognized. During the secondary phase from 5 to 240 minutes the cortical surface PO2 fell by only an additional 23% of the steady state. Even so, when cortical surface PO2 was maintained at this critically low level, the earliest cerebral cortical edema was evident 180 minutes after MCA occlusion. Thereafter, progressive accumulation of edema fluid in the cortex (90 to 170.8 microliters per g of tissue) and in the white matter (19 to 46.2 microliter per g of tissue) was detected by the end of 240 minutes of cerebral ischemia.

Computed tomography in a primate stroke model using selective balloon catheter arterial occlusion.

Ten heparinized nonhuman primates (rhesus monkeys and Papio maryumaya baboons) were subjected to temporary occlusion of an insular branch of the middle cerebral artery with a balloon catheter. An infarct was created. The lesions were monitored in vivo by computed tomography performed at intervals for 1 year. Animals were sacrificed and the lesions examined histologically. The technique obviates the disadvantages of surgical reaction, intravascular thrombosis, and gross cerebral edema observed in prior animal stroke models.

FAILURE OF PROLONGED HYPOCAPNIA, HYPOTHERMIA, OR HYPERTENSION TO FAVORABLY ALTER ACUTE STROKE IN PRIMATES

The effects of induced hypocapnia, hypothermia, and hypertension were surveyed in a primate model of acute stroke during and following a 48-hour period of intensive care. The results were compared to a group of nine control animals previously studied. Hypocapnia (PaCO2=25 torr) was examined in five animals and did not appear to alter the expected mortality, degree of neurological deficit, or frequency of infarction. There was, however, a suggestion that the size of infarction may be reduced. Hypothermia (29 degrees C) in five animals had a detrimental effect in that no animals survived following the intensive care period and all had infarction with massive edema. We speculate that hypothermia caused a sufficient increase in blood viscosity as to compromise collateral flow, thereby accounting for this detrimental effect. Induced hypertension (to 20% above control levels) was abandoned after three animals because of severe systemic effects (cardiac failure and pulmonary edema) resulting in death during the period of intensive care.

Failure of prolonged hypocapnia, hypothermia, or hypertension to favorably alter acute stroke in primates.

The effects of induced hypocapnia, hypothermia, and hypertension were surveyed in a primate model of acute stroke during and following a 48-hour period of intensive care. The results were compared to a group of nine control animals previously studied. Hypocapnia (PaCO2=25 torr) was examined in five animals and did not appear to alter the expected mortality, degree of neurological deficit, or frequency of infarction. There was, however, a suggestion that the size of infarction may be reduced. Hypothermia (29 degrees C) in five animals had a detrimental effect in that no animals survived following the intensive care period and all had infarction with massive edema. We speculate that hypothermia caused a sufficient increase in blood viscosity as to compromise collateral flow, thereby accounting for this detrimental effect. Induced hypertension (to 20% above control levels) was abandoned after three animals because of severe systemic effects (cardiac failure and pulmonary edema) resulting in death during the period of intensive care.

Barbiturate attenuation of the clinical course and pathologic lesions in a primate stroke model.

To evaluate the potential for clinical application, the reputed protective action of barbiturates in cerebral ischemia was tested in a controlled study after segmental middle cerebral artery occlusion in primates. Surviving treated animals promptly recovered consciousness, locomotion, and feeding behavior despite persistent hemiplegia, while control animals ran an indolent course, with slow recovery of poor quality. Cerebral lesions in treated animals were confined to the deep hemispheric structures, while control specimens showed larger deep lesions confluent with extensive areas of cortical infarction. These results are less dramatic than those reported by others, but the protective effect observed in fields of collateral circulation deserves further exploration as an adjunct to medical and surgical management.