Min Of Complete Cerebral Ischemia Research Articles

The effect of glucose upon restitution after transient cerebral ischemia: A summary

The study describes a reproducible model of complete brain ischemia in rats. Rats with different plasma glucose concentrations were exposed to 10 min of complete cerebral ischemia achieved by compression of neck vessels by a pneumatic cuff. All 30 rats, except one, in which pre-ischemic plasma glucose level were lower than 22 mM (range 1.6-22) survived 10 min complete ischemia and made a similar recovery. Ten rats with pre-ischemic plasma glucose levels above 22 mM (range 22-47.2) died from seizures in the post-ischemic period. Post-ischemic treatment of seizures and hyperglycemia in the hyperglycemic rats significantly improved recovery. In conclusion, pre-ischemic hyperglycemia above 22 mM impairs recovery after complete ischemia by inducing seizures, post-ischemic hyperglycemia and lactic acidosis.

Improvement of restitution from cerebral ischemia in hyperglycemic rats by pentobarbital or diazepam.

Hyperglycemic rats were exposed to 10 min of complete cerebral ischemia. Rats with 26 mM (range 23-27 mM) plasma glucose levels died. Rats with the same degree of hyperglycemia, treated after ischemia with pentobarbital (75-95 mg/kg) or diazepam (10-13 mg/kg), survived the observation period of 21 days and recovered, albeit incompletely. The same treatment given to rats with higher degrees of hyperglycemia did not prevent death. It is concluded that postischemic treatment with anticonvulsive drugs may improve clinical restitution from ischemia in moderately hyperglycemic rats.

The arrest of biological time as a bridge to engineered negligible senescence.

Biological systems can remain unchanged for several hundred years at cryogenic temperatures. In several hundred years, current rapid scientific and technical progress should lead to the ability to reverse any biological damage whose reversal is not forbidden by physical law. We therefore explore whether contemporary people facing terminal conditions might be preserved well enough today for their eventual recovery to be compatible with physical law. The ultrastructure of the brain can now be excellently preserved by vitrification, and solutions needed for vitrification can now be distributed through organs with retention of organ viability after transplantation. Current law requires a few minutes of cardiac arrest before cryopreservation of terminal patients, but dogs and cats have recovered excellent brain function after 16-60 min of complete cerebral ischemia. The arrest of biological time as a bridge to engineered negligible senescence, therefore, appears consistent with current scientific and medical knowledge.

ATPases of synaptic plasma membranes from hippocampus after ischemia and recovery during ageing.

Plasticity and relationships between individual ATPases linked to energy-utilizing systems of hippocampus, a very sensitive functional area to both age and ischemia, were studied during ageing on synaptic plasma membranes of 1-year-old "adult" and 2-year-old "aged" rats after 15 min of complete cerebral ischemia and different reperfusion times (01, 24, 48, 72, and 96 h). Activities of Na+, K+, Mg(2+)-ATPase, Mg(2+)-ATPase ouabain insensitive, Na+, K(+)-ATPase, "direct" or "basal" Mg(2+)-ATPase, and acetylcholinesterase (AChE) were evaluated in synaptic plasma membranes, where they play the major role in the regulation of presynaptic nerve ending homeostasis. This in vivo study of recovery time-course from 15 mins of cerebral ischemia indicated specific biochemical assessments of functional meaning: (a) Na+K(+)-ATPase of synaptic plasma membranes in adult and aged animals is stimulated by ischemia; (b) this "hyperactivity" is more markedly related to adult than to aged animals; (c) these abnormalities still persist after 72 and 96 h during the recirculation times, indicating the delayed postischemic suffering of the brain; (d) specific Mg(2+)-ATPase enzyme system possess a lower catalytic power in aged animals than in adult ones, but remained unaltered in adult animals by ischemia and reperfusion; (e) Mg(2+)-ATPase is stimulated in aged animals by ischemia, further increasing during reperfusion up to 72-96 h, indicating the delayed hyperactivity of hippocampus; (f) the increased metabolic activity of hippocampus is indicated by the increased activity of cholinergic system; (g) integrity of synaptic plasma membranes seems not to be altered by 15 min ischemia to a critical extent to compromise their catalytic functionality during reperfusion; (h) AChE activity increases in both adult and aged at some survival times. There are logical reasons for the hypothesis that the modifications in ATPase's catalytic activities in synaptic plasma membranes, which have been modified by ischemia in presynaptic terminals, may play important functional role during recovery time in cerebral tissue in vivo, especially as regards its responsiveness to noxious stimuli, particularly during the recirculation period from acute (or chronic) brain injury.

Free radicals are formed in the brain of fetal sheep during reperfusion after cerebral ischemia.

Free radical production in the brain of acutely anesthetized, exteriorized lamb fetuses (n = 11, gestational age = 135 d) was measured using spin trap methodology. Communications between the vertebral and carotid circulations were tied, producing a two-vessel supply to the brain. Flow probes and occlusion slings were placed around each carotid. The spin trap 2-ethyl-3-hydroxy-2,4,4-trimethyloxazolidine (OXANOH) was infused intermittently into one carotid at a constant rate, and blood samples were taken at intervals from the sagittal sinus. These samples were analyzed for the stable radical OXANO. using electron spin resonance spectrometry. Six animals were subjected to 30 min of complete cerebral ischemia, and five fetuses served as sham-operated control animals. During postischemic reperfusion radical formation increased 2-fold during the first 20 min. However, the elevation of OXANO. in the venous effluent from the brain did not start until the transient hyperemia had passed. It is thus concluded that the increase of OXANO. observed is caused by an augmentation of free radical production during reperfusion. Because the spin trap agent was infused directly into the arterial supply and recovered directly from the venous effluent of the brain, the site of production could be the brain tissue, the endothelial cells of the cerebral circulation, and activated leukocytes. This is the first demonstration of increased radical production from the fetal brain. It is noteworthy that it takes place despite oxygen tension of the reperfusing blood of only 3-3.5 kPa.

Hypoglycemia prevents increase in lactic acidosis during reperfusion after temporary cerebral ischemia in rats.

Sequential 31P and 1H MRS was used to measure cerebral phosphate metabolites, intracellular pH, and lactate in normoglycemic and hypoglycemic rats during 30 min of complete cerebral ischemia and 5.5 h of reperfusion. These results were correlated with brain levels of free fatty acids (FFAs), excitatory amino acids, cations, and water content at death. The lactate/N-acetyl aspartate ratio was not significantly different between groups before or during occlusion. During reperfusion, the ratio was higher in normoglycemic rats from 3 to 85 min (p < or = 0.05), and recovery time was faster in hypoglycemic rats (29 vs 45 min; p = 0.04), suggesting reduced lactate production and faster recovery of aerobic metabolism. During occlusion, significant but comparable decrease of intracellular pH occurred in each group. Intracellular pH was higher in hypoglycemic rats at 140 min and 260 min of reperfusion. Water content, Na and K+ concentrations, and FFA and excitatory amino acid levels were not significantly different between groups, but hypoglycemic rats had less depletion of levels of Mg2+ (p = 0.011). These results show that hypoglycemia has a limited but potentially beneficial effect on postischemic lactic acidosis.

Relationship between extracellular neurotransmitter amino acids and energy metabolism during cerebral ischemia in rats monitored by microdialysis and in vivo magnetic resonance spectroscopy

The time-course of changes in extracellular glutamate and energy metabolism during 30 or 60 min of complete cerebral ischemia and 60–90 min of reperfusion was investigated by microdialysis and magnetic resonance spectroscopy in parallel groups of rats. During the first 10 min of ischemia, adenosine triphosphate (ATP) was completely depleted, and lactate increased 10-fold; after 30 min, intracellular pH had decreased to 6.33 ± 0.11. ATP and lactate did not change further between 30 and 60 min of ischemia. Glutamate increased 30-fold between 10 and 30 min of ischemia and continued to increase in the 60-min ischemia group. After 30 min of reperfusion, glutamate had returned to pre-ischemic levels in both groups. The cellular energy state recovered within 50–60 min after 30 min of ischemia but never returned to more than 60% of baseline values after 60 min of ischemia. The continued increase in extracellular glutamate after total depletion of ATP suggests that glutamate release during ischemia is not entirely energy dependent. Ca 2+-independent glutamate release and failure of energy-dependent glutamate re-uptake mechanisms may result in continued increase in extracellular glutamate. The rapid normalization of extracellular glutamate after 30 and 60 min of ischemia despite differences in the recovery of energy metabolism suggests that the glutamate levels were reduced by an energy-independent mechanism, such as diffusion into the restored circulation.

Effect of dichloroacetate on recovery of brain lactate, phosphorus energy metabolites, and glutamate during reperfusion after complete cerebral ischemia in rats.

The effects of dichloroacetate (DCA) on brain lactate, intracellular pH (pHi), phosphocreatine (PCr), and ATP during 60 min of complete cerebral ischemia and 2 h of reperfusion were investigated in rats by in vivo 1H and 31P magnetic resonance spectroscopy; brain lactate, water content, cations, and amino acids were measured in vitro after reperfusion. DCA, 100 mg/kg, or saline was infused before or immediately after the ischemic period. Preischemic treatment with DCA did not affect brain lactate or pHi during ischemia, but reduced lactate and increased pHi after 30 min of reperfusion (p < 0.05 vs. controls) and facilitated the recovery of PCr and ATP during reperfusion. Postischemic DCA treatment also reduced brain lactate and increased pHi during reperfusion compared with controls (p < 0.05), but had little effect on PCr, ATP, or Pi during reperfusion. After 30 min of reperfusion, serum lactate was 67% lower in the postischemic DCA group than in controls (p < 0.05). The brain lactate level in vitro was 46% lower in the postischemic DCA group than in controls (p < 0.05). DCA did not affect water content or cation concentrations in either group, but it increased brain glutamate by 40% in the preischemic treatment group (p < 0.05). The potential therapeutic effects of DCA on brain injury after complete ischemia may be mediated by reduced excitotoxin release related to decreased lactic acidosis during reperfusion.

Silver impregnability of ischemia-sensitive neocortical neurons after 15 minutes of cardiac arrest in the dog.

The development of postischemic neuronal argyrophilia and the subsequent fate of argyrophilic neurons were studied in dogs after 15 min of complete cerebral ischemia and survival varying from 1 h to 7 days. Histopathological examination of the vulnerable neocortical region was performed using the Nauta degeneration method, and the time course of cellular changes was described. Clear-cut neuronal argyrophilia was found to precede cell body shrinkage and gradual disintegration corresponding to selective neuronal death. To clarify this initial stage of neuronal impregnability, the samples from the animals surviving 8 h postarrest were stained with toluidine blue or processed for electron microscopy, and the distribution of argyrophilic cells was confirmed to be identical with that of hyperchromatic or electron-dense cells. On the other hand, infrequently observed "tissue infarctions" exhibited no silver affinity in spite of apparent cellular damage. These findings indicate that enhanced impregnability is related to cytochemical processes incidental to the phenomenon of "selective neuronal death", which can be readily detected by the Nauta method.

Calcium accumulation by glutamate receptor activation is involved in hippocampal cell damage after ischemia.

Rats exposed to 10 min of complete cerebral ischemia develop necrosis of the CA-1 region of the hippocampus after 2-3 days. We studied the involvement of synaptic transmission for this process by ablation of the afferent input (which is mainly glutamatergic) to CA1 by bilateral destruction of CA-3 neurons (Schafferotomi). The deafferentiation completely prevented the ischemic nerve cell destruction as revealed by histological studies after 6 days. The role of intracellular Ca++ overload was assessed by measurement of the interstitial Ca++ concentration. In control animals the interstitial Ca++ concentration decreases abruptly to 10% of the initial value 1.6 min after the onset of ischemia. The denervated hippocampi, however, showed no decrease during the 10 min of ischemia and hippocampi injected with 2-amino-5-phosphovalerate (APV), a competitive antagonist of the glutamate N-methyl-D-aspartate (NMDA) receptors, displayed a significantly reduced decrease (45% of the initial value) during ischemia. It is concluded that calcium influx via the glutamate-operated channels during the ischemic period is an important link in the development of ischemic brain cell damage.

Cerebral ischemia and reperfusion: prevention of brain mitochondrial injury by lidoflazine.

Mitochondrial degradation is implicated in the irreversible cell damage that can occur during cerebral ischemia and reperfusion. In this study, the effects of 10 min of ventricular fibrillation and 100 min of spontaneous circulation on brain mitochondrial function was studied in dogs in the absence and presence of pretreatment with the Ca2+ antagonist lidoflazine. Twenty-three beagles were separated into four experimental groups: (i) nonischemic controls (ii) those undergoing 10-min ventricular fibrillation, (iii) those undergoing 10-min ventricular fibrillation pretreated with 1 mg/kg lidoflazine i.v., and (iv) those undergoing 10-min ventricular fibrillation followed by spontaneous circulation for 100 min. Brain mitochondria were isolated and tested for their ability to respire and accumulate calcium in a physiological test medium. There was a 35% decrease in the rate of phosphorylating respiration (ATP production) following 10 min of complete cerebral ischemia. Those animals pretreated with lidoflazine showed significantly less decline in phosphorylating respiration (16%) when compared with nontreated dogs. Resting and uncoupled respiration also declined following 10 min of fibrillatory arrest. One hundred minutes of spontaneous circulation following 10 min of ventricular fibrillation and 3 min of open-chest cardiac massage provided complete recovery of normal mitochondrial respiration. Energy-dependent Ca2+ accumulation by isolated brain mitochondria was unimpaired by 10 min of complete cerebral ischemia. However, by 100 min after resuscitation, there was a small, but significant rise in the capacity for mitochondrial Ca2+ sequestration when compared to either control or fibrillated groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Lidoflazine does not improve neurologic outcome when administered after complete cerebral ischemia in primates.

In order to investigate the effects of the calcium entry blocker lidoflazine on neurologic outcome in primates following an episode of global brain ischemia, 12 pigtail monkeys (Macaca nemestrina) were subjected to 17 min of complete cerebral ischemia, followed by 48 h of intensive care treatment and daily neurologic evaluations for 96 h. The monkeys were randomly assigned to receive, in a blind fashion, either lidoflazine 1.0 mg/kg (n = 6) or inactive lidoflazine solvent (n = 6) at 5 min, 8 h, and 16 h postischemia. One monkey in the lidoflazine group did not meet preestablished protocol criteria and was excluded from data analysis. The remaining monkeys were well matched for age, sex, and other physiologic variables. Neurologic outcome was not significantly different between the lidoflazine- and placebo-treated groups (p greater than 0.5). No monkey in either group achieved a normal neurologic exam by 96 h postischemia. Three lidoflazine-treated monkeys and two placebo-treated monkeys died prior to the 96-h neurologic evaluation. These deaths were judged to be neurologic in origin. The authors concluded that lidoflazine does not improve neurologic outcome in primates when administered after 17 min of complete cerebral ischemia.

Effect of lidoflazine on cerebral blood flow and neurologic outcome when administered after complete cerebral ischemia in dogs.

This study tested the hypothesis that lidoflazine, a calcium entry blocking drug, will improve post-ischemic neurologic outcome when administered after a period of complete cerebral ischemia, and that the improvement in outcome is related to an increase in post-ischemic cerebral blood flow (CBF). Complete cerebral ischemia was produced in 31 dogs by temporary ligation of the aorta and venae cavae. In six dogs, 10 min of complete cerebral ischemia was followed by an infusion of lidoflazine 1.0 mg X kg-1 iv over 10 min. CBF and cerebral metabolic rate for oxygen (CMRO2) were measured pre-ischemia, and for 90 min post-ischemia. The results from these six dogs were compared with results previously obtained from eight untreated dogs under similar, but not identical, conditions. The CBF measured post-ischemia in the dogs administered lidoflazine did not differ from the CBF measured post-ischemia in the untreated dogs. Both groups showed an initial hyperemia post-ischemia, followed by significant decreases in CBF from control values by 35 min post-ischemia. The post-ischemic CMRO2 also did not differ between lidoflazine treated and untreated groups. In 25 dogs, 11 min of complete cerebral ischemia was followed by an iv infusion of either lidoflazine 1.0 mg X kg-1 or saline placebo. The same iv infusions were repeated at 8 and 16 h post-ischemia. Seven dogs were excluded from data analysis for failure to meet pre-established protocol criteria. Neurologic injury was evaluated in the remaining dogs at 48 h post-ischemia by an observer blinded to the treatment groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Nicardipine increases cerebral blood flow but does not improve neurologic recovery in a canine model of complete cerebral ischemia.

The effects of the calcium entry blocker nicardipine on CBF, CMRO2, and neurologic outcome following 10 min of complete cerebral ischemia were examined in dogs. In CBF and CMRO2 studies, the CBF in the untreated group (seven dogs) and the nicardipine group (seven dogs; 20 micrograms kg-1 at 30 min postischemia and a subsequent infusion of 2 micrograms kg-1 min-1 for 90 min) initially increased to 300-400% and then returned to preischemic values at 30 min postischemia. Thereafter the CBF in the untreated group significantly decreased to 50% of preischemic values for the following 90-min period (hypoperfusion), while the CBF in the nicardipine group did not differ from preischemic values. The CMRO2 in both groups decreased to approximately 50-80% of preischemic values after 15 min postischemia and did not differ between the groups throughout the study. In neurologic outcome studies, 18 dogs were divided into three groups (of six dogs each): untreated; saline infusion only, posttreated; nicardipine as in CBF and CMRO2 studies, pretreated; nicardipine 20 micrograms kg-1 at 2 min preischemia and a subsequent infusion of 2 micrograms kg-1 min-1 from immediately postischemia to 120 min postischemia. Nicardipine treatment initiated either before or after ischemia failed to improve neurologic outcome at 48 h postischemia. Thus, the increase of postischemic global CBF by nicardipine is not accompanied by neurologic recovery in a canine model of complete cerebral ischemia.

Effects of mannitol on cerebral circulation after transient complete cerebral ischemia in dogs.

The effects of hypertonic mannitol on postischemic cerebral circulation were studied in 20 dogs. Mannitol, 2 g/kg iv, was infused into ten dogs during a 2-h period starting 1.5 h after 6 min of complete cerebral ischemia. One hour postischemia, regional cerebral blood flow (rCBF) was 36% in the control group (ten dogs) and 39% in the mannitol group. In the control group, rCBF increased gradually thereafter to 70% of the preischemic value 10 h postischemia, whereas the restoration of rCBF was suppressed in the mannitol group. During the postischemic period, intracranial pressure (ICP) increased significantly in the control group, but it did not change significantly in the mannitol group. The effects of mannitol on rCBF and ICP in the present study can be explained primarily by decreased body water due to urinary loss. The administration of mannitol does not necessarily improve postischemic cerebral circulation.

Glycerol as an indicator of lipid degradation in bicuculline-induced seizures and experimental cerebral ischemia.

Glycerol, the end product of phospholipid degradation, was measured in cat brains under pathophysiological conditions known to cause activation of lipolysis, namely, bicuculline-induced seizures, permanent focal cerebral ischemia (2 hr of middle cerebral artery occlusion), and global cerebral ischemia (15 min of complete cerebral ischemia with or without 2 hr of recirculation). In addition, ATP and lactate were measured in order to correlate the activation of lipid degradation with disturbances in the energy-producing metabolism. A highly significant increase in the tissue glycerol content was observed after 1 hr of bicuculline-induced seizures (from 0.29 +/- 0.07 mumol/g in control animals to 1.30 +/- 0.06 mumol/g in seizure animals; P less than 0.001) or after 15 min of complete cerebral ischemia (from 0.29 +/- 0.07 to 1.17 +/- 0.14 mumol/g; P less than 0.01). Furthermore, a close correlation was found between the increase in glycerol and the increase in lactate or decrease in ATP after permanent focal ischemia. In contrast, following recirculation after complete cerebral ischemia, restoration of the energy pool did not lead to a reduction of the glycerol formed during ischemia. It is concluded that glycerol is a useful indicator of lipid degradation under pathological conditions. Since glycerol formed during vascular occlusion is trapped in brain cells, presumably owing to low glycerol kinase activity, it can be used as a stable postischemic indicator of ischemia-induced lipid degradation.

Nimodipine improves outcome when given after complete cerebral ischemia in primates.

Twenty-seven pigtailed monkeys (Macaca nemestrina) were subjected to 17 min of complete cerebral ischemia followed by 96 h of intensive care treatment. Fourteen of the monkeys were assigned randomly to the treatment group and received nimodipine 10 micrograms . kg-1 5 min postischemia followed by 1 microgram . kg-1 . min-1 for 10 h. Six monkeys (three treated) failed to meet preestablished protocol criteria and were excluded. The remaining treated and untreated monkeys were well matched for age, sex, and other physiologic variables. Neurologic outcome at 96 h postischemia was significantly better in the nimodipine-treated monkeys than in the controls. Eight of the 11 treated animals had an apparent normal level of consciousness; four of these had no detectable neurologic deficits and a fifth had only a slight motor apraxia. Only two of the 10 untreated animals had an apparent normal level of consciousness, and all had major neurologic deficits. Histopathologic examination showed variable ischemic neuronal change and infarction to involve gray matter in distal arterial perfusion zones. Significant white matter changes were not observed. A histopathologic scoring system yielded a significantly better mean score for the treated group than for the untreated group, and there was significant correlation between neurologic function and histopathologic findings. The authors conclude that nimodipine improves the neurologic outcome when given after an episode of complete cerebral ischemia in primates, and they recommend controlled clinical trials in patients resuscitated after cardiac arrest.

Complete cerebral ischemia. An ultrastructural study.

Neuronal, astrocytic, and oligodendrocytic elements in several brain loci of the cat were examined at the light and electron microscopic level immediately after periods of complete cerebral ischemia (CCI) uncomplicated by post-ischemic recirculation. Such CCI episodes ranged from 1.5–25 min duration and were methodically produced in a cat model employing rigorous physiological controls. Subsequent to these CCI insults, morphological alterations occurred in a homogeneous manner within each cell type of all loci examined; however, variation in the temporal onset and magnitude of alterations among the various cell types was observed. With brief ischemic insults all cell nuclei demonstrated pronounced nuclear alterations, while their cytoplasmic organelles displayed minimal change. Chromatin clumping and nucleolar condensation were observed in both neurons and glia subsequent to 1.5–5 min of CCI, respectively. With increasing durations of CCI such changes were more dramatic and conspicuous alterations of the cytoplasmic organelles were observed. On the basis of extensive morphological analyses the present study illustrates that nuclear alterations are the first to occur subsequent to CCI. The homogeneity of neuronal involvement seen subsequent to CCI uncomplicated by post-ischemic recirculation is inconsistent with the “selective vulnerability” purported to occur by others. The significance of this inconsistency remains to be assessed; yet, the suggestion is advanced that post-ischemic recirculation may be a factor in the genesis of such vulnerability.