bluuebunny/crossref_metadata_2025 huggingface dataset:
The Use of In Vivo Fluorescence Image Sequences to Indicate the Occurrence and Propagation of Transient Focal Depolarizations in Cerebral Ischemia
Authors: Strong A. J. , Harland S. P. , Meldrum B. S. , Whittington D. J.
Published: November 2003

A method for the detection and tracking of propagated fluorescence transients as indicators of depolarizations in focal cerebral ischemia is described, together with initial results indicating the potential of the method. The cortex of the right cerebral hemisphere was exposed for nonrecovery experiments in five cats anesthetized with chloralose and subjected to permanent middle cerebral artery (MCA) occlusion. Fluorescence with 370-nm excitation (attributed to the degree of reduction of the NAD/H couple) was imaged with an intensified charge-coupled device camera and digitized. Sequences of images representing changes in gray level from a baseline image were examined, together with the time courses of mean gray levels in specified regions of interest. Spontaneous increases in fluorescence occurred, starting most commonly at the edge of areas of core ischemia; they propagated usually throughout the periinfarct zone and resolved to varying degrees and at varying rates, depending on proximity of the locus to the MCA input. When a fluorescence transient reached the anterior cerebral artery territory, its initial polarity reversed from an increase to a decrease in fluorescence. An initial increase in fluorescence in response to the arrival of a transient may characterize cortex that will become infarcted, if pathophysiological changes in the periinfarct zone are allowed to evolve naturally.
Inhibition of Tumor Necrosis Factor Alpha (TNFα) Activity in Rat Brain is Associated with Cerebroprotection after Closed Head Injury
Authors: Shohami E. , Bass R. , Wallach D. , Yamin A. , Gallily R.
Published: November 2003

We recently demonstrated that closed head injury (CHI) in the rat triggers the production of tumor necrosis factor alpha (TNFα) in the contused hemisphere. Other investigations have shown that this cytokine plays a role in the inflammatory response following trauma. The present study was designed to determine whether inhibition of TNFα production or activity affects the development of cerebral edema as well as neurological dysfunction and hippocampal cell loss after CHI. To this end, we used two pharmacological agents, each acting via a different mechanism: pentoxifylline (PTX), which attenuates the production of TNFα, and tumor necrosis factor binding protein (TBP), a physiological inhibitor of TNFα activity. Both agents significantly lessened peak edema formation at 24 h and facilitated the recovery of motor function for ≤14 days postinjury. In addition, TBP attenuated disruption of the blood-brain barrier and protected hippocampal cells. PTX significantly lowered the brain TNFα level (by ∼80%), and TBP completely abolished the activity of recombinant human TNF when they were added at the same time in the in vitro bioassay. We suggest, therefore, that a decrease in TNFα level or the inhibition of its activity is accompanied by reduced brain damage.
Hypertonic Environment Prevents Depolarization and Improves Functional Recovery from Hypoxia in Hippocampal Slices
Authors: Huang Rong , Aitken Peter G. , Somjen George G.
Published: November 2003

Treatments that postpone hypoxic spreading depression (SD)-like depolarization (also called anoxic depolarization) facilitate recovery of function after transient cerebral hypoxia. Hypertonia reduces cerebral excitability, and we tested whether it also offers protection against SD-like depolarization and hypoxia. Oxygen was withdrawn from hippocampal slices bathed in normal artificial cerebrospinal fluid (ACSF) and, simultaneously, from slices cut from the same hippocampus but bathed in strongly hypertonic ACSF. Extracellular osmolarity (πo) was increased by adding 100 m M mannitol or fructose to ACSF. Slices in normal πo underwent SD-like negative extracellular voltage shift (ΔVo). The hypertonic slices usually showed no SD-like ΔVo but only a small, gradual negative voltage shift. Hypertonia also prevented the precipitate drop of interstitial calcium level ([Ca2+]o). When oxygenation and normal osmolarity were restored, synaptic transmission in the previously hypertonic slices recovered completely, but 3 h after reoxygenation orthodromically transmitted population spikes of the control slices recovered only 25.1% of the initial control amplitude. We conclude that hypertonic treatment during hypoxia improves subsequent recovery of synaptic function. The protection is probably due to the prevention of calcium uptake by blocking the SD-like depolarization, with the prevention of hypoxic cell swelling playing a lesser role.
Neuroprotection after Several Days of Mild, Drug-Induced Hypothermia
Authors: Nurse Suzanne , Corbett Dale
Published: November 2003

Stroke trials are initiated after demonstrated pharmacological protection in animal models. NBQX protects CA1 neurons against global ischemia; however, this glutamate antagonist induces a period of subnormal temperature (e.g., a decrease of only 1.0–1.5°C) lasting several days. In this study, NBQX (3 × 30 mg/kg, i.p.) was administered starting 60 min after reperfusion, and brain temperature had declined significantly below vehicle-treated animals by 2 h after reperfusion. When the postischemic brain temperature of NBQX-treated gerbils was regulated, no neuronal protection was found. Mimicking an NBQX-induced temperature profile for 28 h postischemia yielded histological protection 4 days later comparable to that of NBQX. However, both the NBQX and temperature simulation groups showed decreased protection after 10-day survival. Our data suggest that a protracted period of subnormal temperature during the postischemic period can obscure the interpretation of preclinical drug studies.
Color Duplex Measurement of Cerebral Blood Flow Volume: Intra- and Interobserver Reproducibility and Habituation to Serial Measurements in Normal Subjects
Authors: Schöning Martin , Scheel Peter
Published: November 2003

Color duplex flowmetry of internal carotid and vertebral arteries permits estimation of intravascular flow volumes and global cerebral blood flow volume (CBFV) by summing the flow volumes measured in each of the four extracranial vessels. Intravascular flow volumes were calculated as the product of angle-corrected time-averaged flow velocity and the cross-sectional area of the vessel. The reliability of this new method was tested in a prospective, intra- and interdiane, intra- and interobserver reproducibility study of 32 healthy subjects aged 7–57 years. In each subject, CBFV was tested by each observer twice on day 1 and once on day 2 in consecutive recordings. In each artery, both examiners found closely similar mean intravascular flow volumes. Intradiane interobserver reproducibility of CBFV was high on both days (correlation coefficient, CC, 0.90 and 0.85, p ≤ 0.0001; coefficient of variance, CV, 10.0 and 10.4%, respectively), as was the interdiane comparison (CC = 0.81, p ≤ 0.0001; CV ≤ 13.3%). Intraobserver reproducibility was even higher. On both days, there was a progressive decrease in CBFV from each subject's first to the last examination within a 1-h examination period (day 1: 717 ± 150 ml/min to 690 ± 120 ml/min; difference, p ≤ 0.05; day 2: 700 ± 120 ml/min to 665 ± 126 ml/min; difference, p ≤ 0.01). This habituation effect was more pronounced in subjects with high initial CBFV. Reproducibility of CBFV is comparable to that of mean CBF measurements with 133Xe inhalation and H215O positron emission tomography techniques reported by other groups. This method makes serial bedside-monitoring of CBFV feasible without posing the risks of radiation exposure.
Intracisternal Basic Fibroblast Growth Factor (bFGF) Enhances Behavioral Recovery following Focal Cerebral Infarction in the Rat
Authors: Kawamata Takakazu , Alexis Nancy E. , Dietrich W. Dalton , Finklestein Seth P.
Published: November 2003

Basic fibroblast growth factor (bFGF) is a potent neurotrophic agent that promotes neuronal survival and outgrowth. Previous studies have shown that bFGF, administered intraventricularly or intravenously before or within hours after ischemia, reduces infarct size and neurological deficits in models of focal cerebral ischemia in rats. In the current study, we tested the hypothesis that bFGF, administered at later time points after ischemia, might improve behavioral recovery without affecting infarct size. Mature Sprague–Dawley rats received bFGF (1 μg/injection) or vehicle by biweekly intracisternal injection for 4 weeks, starting at 1 day following permanent proximal middle cerebral artery (MCA) occlusion. Animals were examined every other day using four different behavioral tests to assess sensorimotor and reflex function. At 4 weeks after ischemia, there was no difference in infarct volume between bFGF- and vehicle-treated animals. There was, however, an enhancement in the rate and degree of behavioral recovery among bFGF-treated animals, as measured by all four tests. There were no apparent side effects of bFGF treatment, except that bFGF-treated animals tended to recover body weight more slowly than did vehicle-treated animals following stroke. The mechanisms of enhancement of behavioral recovery by bFGF require further study, but may include protection against retrograde neuronal death and/or stimulation of neuronal sprouting.
Global Ischemia Induces Immediate-Early Genes Encoding Zinc Finger Transcription Factors
Authors: Honkaniemi Jari , Sharp Frank R.
Published: November 2003

Ischemia induces immediate-early genes (IEGs) in brain. Since prolonged expression of some IEGs may precede neuronal death, some researchers have suggested that these IEGs mediate neuronal death. We therefore examined the effect of 5 and 10 min of global ischemia on the expression of the IEGs NGFI-A, NGFI-B, NGFI-C, egr-2, egr-3, and Nurr1 in gerbil brain. All of the IEGs were induced after 30 min of reperfusion in the hippocampus. Most of them were induced in several other regions as well, including cortex, hypothalamus, thalamus, and amygdala. The acute IEG induction decreased in most brain areas by 2–6 h. However, at 24 h following 5 min of ischemia NGFI-A continued to be expressed in the CA1 region and dentate gyrus. In the dentate gyrus, NGFI-C continued to be expressed for 24 h and egr-3 for as long as 72 h. In other brain areas, all of the IEGs returned to control levels by 72 h except in CA1, where most messenger RNA (mRNA) levels were decreased; this decrease correlated with marked neuronal loss. The persistent expression of NGFI-A in CA1 neurons destined to die and the persistent expression of NGFI-A, NGFI-C, and egr-3 genes in dentate granule cell neurons that survive may indicate that some transcription factors modulate cell death whereas others support cell survival when expressed for prolonged periods. The protein products of several transcription factors, including c- fos, are known to downregulate their own expression. The persistent expression of NGFI-A in the CA1 neurons destined to die could therefore be due to ischemia-induced transcriptional activation caused by, e.g., increased intracellular calcium levels plus a lack of negative feedback caused by the blockade of the translation of NGFI-A mRNA into protein.
L-Arginine Does Not Improve Cortical Perfusion or Histopathological Outcome in Spontaneously Hypertensive Rats Subjected to Distal Middle Cerebral Artery Photothrombotic Occlusion
Authors: Prado Ricardo , Watson Brant D. , Zhao Weizhao , Yao Hiroshi , Busto Raul , Dietrich W. Dalton , Ginsberg Myron D.
Published: November 2003

The potential of nitric oxide (NO) to influence positively or negatively the outcome of mechanically induced focal cerebral ischemia is still controversial. Recent evidence suggests that NO of vascular origin, whether synthesized from exogenously administered L-arginine (L-Arg) or from NO donor compounds, is beneficial but that of neuronal origin is not. However, the therapeutic potential of NO to ameliorate stroke induced by arterial thrombosis has not been reported. We assessed the therapeutic effect of L-Arg administration in spontaneously hypertensive rats (SHR) subjected to permanent photothrombotic occlusion of the distal middle cerebral artery (dMCA). The ipsilateral carotid artery was left unligated to enhance L-Arg delivery into the putative penumbral region. Local CBF (LCBF) was assessed at 30 min by the [14C]iodoantipyrine technique (n = 9), while histological infarct volumes and index of peripheral ischemic cell change were determined at 3 days (n = 7). Rats (n = 9) given 300 mg/kg L-Arg at 18 and 3 h before photothrombotic dMCA occlusion and at 5 min afterward displayed no significant differences in LCBF compared with animals (n = 8) injected with water (the carrier vehicle) and similarly irradiated. Infarct volumes were also similar, being 37.0 ± 9.7 mm3 (SD) in the vehicle-treated and 49.1 ± 17.2 mm3 (SD) in the L-Arg-treated groups (both n = 7), as were assessments of ischemic neuronal density in the penumbra. In contrast, L-Arg administered intravenously in a dose of 300 mg/kg to nonischemic SHR (n = 5) increased cortical CBF by ∼75% during a 70-min observation period. We conclude that thrombotic processes superimposed upon cerebral ischemia may facilitate tissue reactions that offset the potentially beneficial effect of L-Arg, and this caveat must be considered when proposing L-Arg for clinical treatment of focal thrombotic stroke.
Quantitation of Regional Cerebral Blood Flow with 15O-Butanol and Positron Emission Tomography in Humans
Authors: Herzog H. , Seitz R. J. , Teilmann L. , Kops E. Rota , Jülicher F. , Schlaug G. , Kleinschmidt A. , Müller-Gärtner H. W.
Published: November 2003

We describe the implementation and validation of a combined dynamic–autoradiographic approach for measuring the regional cerebral blood flow (rCBF) with 15O-butanol. From arterial blood data sampled at a rate of 1 s and list mode data of the cerebral radioactivity accumulated over 100 s, the time shift between blood and tissue curves, the dispersion constant DC, the partition coefficient p, and the CBF were estimated by least squares fitting. Using the fit results, a pixel-by-pixel parametrization of rCBF was computed for a single 40-s (autoradiographic) 15O-butanol uptake image. The mean global CBF found in 27 healthy subjects was 49 ± 8 ml 100 g−1 min−1. Gray and white matter rCBF were 83 ± 20 and 16 ± 3 ml 100 g−1 min−1, respectively, with a corresponding partition coefficient p of 0.77 ± 0.18 and 0.77 ± 0.29 ml/g in both compartments. The quantitative images resulted in a significantly higher gray matter rCBF than the autoradiographic images.
Increased Expression of Endothelin B Receptor mRNA following Subarachnoid Hemorrhage in Monkeys
Authors: Hino Akihiko , Tokuyama Yoshiharu , Kobayashi Masahiko , Yano Mitsuo , Weir Bryce , Takeda Jun , Wang Xiaoyu , Bell Graeme I. , Macdonald R. Loch
Published: November 2003

These studies tested the hypothesis that the cerebral vasospasm that follows subarachnoid hemorrhage (SAH) is due to alterations in endothelin (ET) and ET receptor expression. Eight monkeys underwent cerebral angiography and induction of SAH. Angiography was repeated 7 days later to confirm the presence of cerebral vasospasm, and animals were killed. RNA was isolated from right (vasospastic) and left (control) side middle cerebral arteries and surrounding cerebral cortex. The levels of prepro (PP) ET-1 (ppET-1) and ppET-3 and ETA and ETB receptor mRNAs were determined using a quantitative reverse transcriptase polymerase chain reaction-based assay. ET-1 peptide was also measured in CSF at baseline and after 7 days. Specific agonist binding to ETA and ETB receptors in both middle cerebral arteries and in surrounding brain cortex was measured in three animals by autoradiographic binding assays. Levels of ETB receptor mRNA were 3.4 ± 2.2-fold higher in the right than in the left cerebral arteries (p < 0.01). There were no significant differences in the levels of ppET-1, ppET-3, or ETA receptor mRNA in cerebral arteries. ET-1 peptide was not elevated in CSF. Levels of ETA and ETB receptor mRNAs were 2.6 ± 1.1- and 2.1 ± 1.3-fold higher, respectively, in the right than in the left cerebral cortex, while the level of ppET-3 mRNA was 2.1 ± 1.0-fold lower. There were no differences in ppET-1 mRNA levels between right and left cerebral cortex. Binding to ETA and ETB receptors in cerebral arteries and cortex did not differ significantly between right and left sides. These results do not support the hypothesis that overexpression of ET-1 is the principal cause of vasospasm, but rather they suggest that SAH causes complex changes in the ET system that together are responsible for the cellular response to SAH.
An Alternative Method for the Quantitation of Neuronal Damage after Experimental Middle Cerebral Artery Occlusion in Rats: Analysis of Behavioral Deficit
Authors: Aronowski Jaroslaw , Samways Emma , Strong Roger , Rhoades Howard M. , Grotta James C.
Published: November 2003

We tested the hypothesis that increasing durations of focal ischemia that have been shown to result in enlargement of cortical infarct will be associated with progression of behavioral dysfunction that can be measured by a battery of tests sufficiently sensitive and reproducible to detect a positive effect of pharmacotherapy. Untreated or N-methyl-D-aspartate receptor antagonist (CNS-1102)-treated spontaneously hypertensive rats underwent 45, 60, 90, or 120 min of tandem middle cerebral and common carotid artery occlusion followed by reperfusion. We then evaluated the extent of damage and its recovery for up to 21 days using nine behavioral tests aimed at analyzing strength, coordination, and bilateral asymmetry. Also using a graded bioassay that employs a curve-fitting computer program (ALLFIT) to correlate duration of ischemia with degree of behavioral dysfunction, we calculated the average maximal behavioral dysfunction and duration of ischemia required to produce half-maximal behavioral dysfunction and compared these values in untreated controls with analogous values obtained from animals treated with CNS-1102. Three behavioral tests, forearm flex, tape (somatosensory neutralization), and foot-fault placing, were each separately and combined able to distinguish between the degrees of damage produced by increasing durations of ischemia. The behavioral abnormalities assessed using the tape test were reversible within a week, whereas those using forearm flex or foot-fault tests persisted for at least 21 days. CNS-1102 significantly reduced behavioral dysfunction measured by all three tests. This analysis of behavioral dysfunction represents a useful experimental model to grade efficacy of therapies aimed at protecting the brain from damage produced by acute stroke and might also be used to assess recovery from preexisting ischemic damage.
Exercise-Induced Changes in Local Cerebral Glucose Utilization in the Rat
Authors: Vissing John , Andersen Martin , Diemer Nils H.
Published: November 2003

In exercise, little is known about local cerebral glucose utilization (LCGU), which is an index of functional neurogenic activity. We measured LCGU in resting and running (≈85% of maximum O2 uptake) rats (n = 7 in both groups) previously equipped with a tail artery catheter. LCGU was measured quantitatively from 2-deoxy-D-[1-14C]glucose autoradiographs. During exercise, total cerebral glucose utilization (TCGU) increased by 38% (p < 0.005). LCGU increased (p < 0.05) in areas involved in motor function (motor cortex 39%, cerebellum ≈110%, basal ganglia ≈30%, substantia nigra ≈37%, and in the following nuclei: subthalamic 47%, posterior hypothalamic 74%, red 61%, ambiguus 43%, pontine 61%), areas involved in sensory function (somatosensory 27%, auditory 32%, and visual cortex 42%, thalamus ≈75%, and in the following nuclei: Darkschewitsch 22%, cochlear 51%, vestibular 30%, superior olive 23%, cuneate 115%), areas involved in autonomic function (dorsal raphe nucleus 30%, and areas in the hypothalamus ≈35%, amygdala ≈35%, and hippocampus 29%), and in white matter of the corpus callosum (36%) and cerebellum (52%). LCGU did not change with exercise in prefrontal and frontal cortex, cingulum, inferior olive, nucleus of solitary tract and median raphe, lateral septal and interpenduncular nuclei, or in areas of the hippocampus, amygdala, and hypothalamus. Glucose utilization did not decrease during exercise in any of the studied cerebral regions. In summary, heavy dynamic exercise increases TCGU and evokes marked differential changes in LCGU. The findings provide clues to the cerebral areas that participate in the large motor, sensory, and autonomic adaptation occurring in exercise.
Induction of Platelet-Derived Growth Factor β-Receptor in Focal Ischemia of Rat Brain
Authors: Iihara Koji , Sasahara Masakiyo , Hashimoto Nobuo , Hazama Fumitada
Published: November 2003

Our previous study on the ischemia-induced expression of platelet-derived growth factor (PDGF)-B chain in the rat brain prompted us to examine expression of PDGF β-receptor in the ischemic brain. Focal ischemia was induced by permanent tandem occlusion of middle cerebral and common carotid arteries in spontaneously hypertensive rats. Northern analysis revealed that ischemia significantly increased expression of the receptor in the ischemic neocortex at 4 and 7 days (328 ± 109%; 323 ± 119%, respectively, over control: n = 4, p < 0.05 versus sham). Neurons in infarct transiently snowed increased immunostaining for the receptor at 1 day, whereas neurons in periinfarct area showed sustained and increased immunoreactivity from 1 to 14 days postischemia. Reactive glial cells in the external capsule and in molecular layer of the neocortex adjacent to infarct possessed enhanced immunoreactivity from 1 to 21 days. Furthermore, marked immunoreactivity was observed on brain macrophages in infarct and on the abluminal side of capillaries surrounding infarct from 4 to 7 days. These results demonstrated that ischemic insult increases expression of the PDGF β-receptor at both the mRNA and protein level in the brain, suggesting its important role in cellular cascade of the ischemic brain.
Nitric Oxide Synthase Inhibition by L-NAME Prevents Brain Acidosis during Focal Cerebral Ischemia in Rabbits
Authors: Regli Luca , Held Mark C. , Anderson Robert E. , Meyer Fredric B.
Published: November 2003

This experiment examined the effects of nitric oxide (NO) synthase inhibition on brain intracellular pH, regional cortical blood flow, and NADH fluorescence before and during 3 h of focal cerebral ischemia using in vivo fluorescence imaging. Thirty fasted rabbits under 1% halothane were divided into four treatment groups receiving Nω-nitro-L-arginine methyl ester (L-NAME) intravenously at 20 min prior to ischemia (0.1, I, and 10 mg/kg and 1 mg/kg + 5 mg/kg L-arginine) and two control groups (nonischemic and ischemic). In ischemic controls, brain pHi declined to 6.73 ± 0.03 at 30 min and remained acidotic through the remainder of the ischemic period. In the 0.1 mg/kg group, brain pHi fell after 30 min of ischemia to 6.76 ± 0.05 ( p < 0.05), but then improved progressively despite occlusion. In the 1 mg/kg group, brain pHi remained normal despite middle cerebral artery (MCA) occlusion. In the 10 mg/kg group and in the combined L-NAME + L-arginine group, pHi fell after 30 min of ischemia to 6.81 ± 0.03 ( p < 0.05) and remained acidotic. During occlusion, regional cortical blood flow dropped in a dose-dependent manner. After 3 h of ischemia, regional cortical blood flow was 33.9 ± 10.9 and 25.1 ± 8.9 ml/100 g/min at doses of 0.1 and 10.0 mg/kg, respectively. L-NAME treatment did not significantly alter the increased NADH fluorescence that accompanied occlusion. This study shows that L-NAME can prevent intracellular brain acidosis during focal cerebral ischemia independent from regional cortical blood flow changes. This experiment suggests that NO is involved in pHi regulation during focal cerebral ischemia.
Neuronal Damage following Intraspinal Injection of a Nitric Oxide Synthase Inhibitor in the Rat
Authors: Yezierski Robert P. , Liu Shanliang , Ruenes Gladys L. , Busto Raul , Dietrich W. Dalton
Published: November 2003

Intraspinal microinjection of the nonspecific nitric oxide synthase (NOS) inhibitor N-nitro-L-arginine methyl ester (L-NAME) was used to determine if inhibition of NOS results in morphological changes in the rat spinal cord. Following spinal injections of 100–750 m M L-NAME (pH 7.0), 1.0–500 m M L-NAME (pH 2.5–5.4), or L-NAME + L-arginine, quantitative analysis of morphological changes revealed a positive dose-response relationship between L-NAME and neuronal loss. This effect was blocked by L-arginine and was inversely related to spinal levels of NOS enzyme activity. Results of this study have shown the importance of basal NOS activity in maintaining the structural integrity of spinal neurons. It is proposed that the effects of L-NAME on nitric oxide (NO) production leads to decreased blood flow, secondary to vasoconstriction, and a hypoxic–ischemic reaction in spinal tissue. The results suggest that a potential contributing factor to neuronal damage in pathological conditions such as spinal cord injury may be the decreased production of nitric oxide.
The Effect of Hyperglycemia on Cerebral Metabolism during Hypoxia-Ischemia in the Immature Rat
Authors: Vannucci Robert C. , Brucklacher Robert M. , Vannucci Susan J.
Published: November 2003

Unlike adults, hyperglycemia with circulating glucose concentrations of 25–35 m M/L protects the immature brain from hypoxic–ischemic damage. To ascertain the effect of hyperglycemia on cerebral oxidative metabolism during the course of hypoxia–ischemia, 7-day postnatal rats underwent unilateral common carotid artery ligation followed by exposure to 8% O2 for 2 h at 37°C. Experimental animals received 0.2 cc s.c. 50% glucose at the onset of hypoxia–ischemia, and 0.15 cc 25% glucose 1 h later to maintain blood glucose concentrations at 20–25 m ML for 2 h. Control rat pups received equivalent concentrations or volumes of either mannitol or 1 N saline at the same intervals. The cerebral metabolic rate for glucose (CMRglc.) increased from 7.1 (control) to 20.2 μmol 100 g−1 min−1 in hyperglycemic rats during the first hour of hypoxia–ischemia, 79 and 35% greater than the rates for saline- and mannitol-injected animals at the same interval, respectively ( p < 0.01). Brain intracellular glucose concentrations were 5.2 and 3.0 m M/kg in the hyperglycemic rat pups at 1 and 2 h of hypoxia–ischemia, respectively; glucose levels were near negligible in mannitol- and saline-treated animals at the same intervals. Brain intracellular lactate concentrations averaged 13.4 and 23.3 m M/kg in hyperglycemic animals at 1 and 2 h of hypoxia–ischemia, respectively, more than twice the concentrations estimated for the saline- and mannitol-treated littermates. Phosphocreatine (PCr) and ATP decreased in all three experimental groups, but were preserved to the greatest extent in hyperglycemic animals. Results indicate that anaerobic glycolytic flux is increased to a greater extent in hyperglycemic immature rats than in normoglycemic littermates subjected to cerebral hypoxia–ischemia, and that the enhanced glycolysis leads to greater intracellular lactate accumulation. Despite cerebral lactosis, energy reserves were better preserved in hyperglycemic animals than in saline-treated controls, thus accounting for the greater resistance of hyperglycemic animals to hypoxic–ischemic brain damage.
Continuous Measurement of Changes in Internal Carotid and Vertebral Arterial Blood Flow with Chronically Implanted Ultrasonic Doppler Probes in Anesthetized and Conscious Rabbits
Authors: Yu Ying-Hui , Zhu B-S , Blessing W. W.
Published: November 2003

Chronically implanted ultrasonic Doppler flowmeters were used to obtain simultaneously recorded flow velocity signals from internal carotid and vertebral arteries, and the sagittal sinus, in rabbits. All three signals increased to 144 ± 7–215 ± 35% of baseline during hypercapnia (arterial Pco2 55 mmHg) in both anesthetized and conscious animals. During the period of change in inspired CO2, the relationship between simultaneously recorded mean internal carotid and mean sagittal sinus signals was linear, with the correlation ranging from 0.83 to 0.96. Since forebrain arterial inflow must approximate forebrain venous outflow, the high correlation between internal carotid and sagittal sinus signals indicates that these measures provide reliable and valid indices of cerebral blood flow (CBF). Vertebral and internal carotid angiography confirmed the location of Doppler probe. Chronically implanted ultrasonic Doppler flowmeters can, thus, provide continuous noninvasive measurements of cerebral arterial flow in both anesthetized and conscious rabbits.
GABAA Receptor Activation Attenuates Excitotoxicity but Exacerbates Oxygen—Glucose Deprivation-Induced Neuronal Injury In Vitro
Authors: Muir Judith K. , Lobner Doug , Monyer Hannelore , Choi Dennis W.
Published: November 2003

We examined the effects of GABA receptor stimulation on the neuronal death induced by exogenously added excitatory amino acids or combined oxygen–glucose deprivation in mouse cortical cell cultures. Death induced by exposure to NMDA, AMPA, or kainate was attenuated by addition of GABA or the GABAA receptor agonist, muscimol, but not by the GABAB receptor agonist, baclofen. The antiexcitotoxic effect of GABAA receptor agonists was blocked by bicuculline or Picrotoxin. In contrast, GABA or muscimol, but not baclofen, markedly increased the neuronal death induced by oxygen–glucose deprivation. Muscimol potentiation of neuronal death was associated with increased glutamate efflux to the bathing medium, and increased cellular 45Ca2+ accumulation; it was blocked by MK-801, but not NBQX, suggesting mediation by NMDA receptors. Bicuculline only weakly attenuated muscimol potentiation of oxygen–glucose deprivation-induced neuronal death, probably because it itself increased this death. Present results raise a note of caution in the proposed use of GABAA receptor stimulation to limit ischemic brain damage in vivo.
Estimation of Cerebral Blood Flow at Bedside by Continuous Jugular Thermodilution
Authors: Mélot Christian , Berré Jacques , Moraine Jean-Jacques , Kahn Robert Jean
Published: November 2003

The Kety-Schmidt technique can be regarded as the reference method for the measurement of cerebral blood flow (CBF). However, the method is somewhat cumbersome for routine use in the intensive care unit (ICU) at the bedside. The continuous thermodilution technique developed many years ago for the measurement of coronary sinus blood flow can be applied for the measurement of jugular blood flow (JBF). However, the measurement of JBF by thermodilution has never been validated using the Kety-Schmidt reference method. We first validate the continuous thermodilution in vitro by comparison with a volumetric flow. The thermodilution method is accurate for flows between 50 and 900 ml min−1 with a mean difference volumetric-thermodilution flow of −1 ± 18 ml min−1 (mean ± SD), and precise with a coefficient of variability ranging between 1.21% and 2.50%. In vivo accuracy was assessed by comparing in 15 comatose patients CBF measured using the Kety-Schmidt (CBFKS) method and estimated from JBF measured by thermodilution (CBFTH) at four levels of arterial PaCO2 (25, 30, 35, and 40 mm Hg). The mean difference CBFKS — CBFTH is −0.9 ± 3.6 ml min−1 100 g−1. In vivo precision of the method was good, with a coefficient of variability of 4.1% in mean. We conclude that jugular continuous thermodilution technique is a reliable method for estimating CBF at the bedside. This technique allows repeated measurements jugular bulb blood sampling for brain metabolic studies.
Kinetic Evaluation of [11C]Dihydrotetrabenazine by Dynamic PET: Measurement of Vesicular Monoamine Transporter
Authors: Koeppe R. A. , Frey K. A. , Vander Borght T. M. , Karlamangla A. , Jewett D. M. , Lee L. C. , Kilbourn M. R. , Kuhl D. E.
Published: November 2003

(+)-α-[11C]Dihydrotetrabenazine (DTBZ) binds to the vesicular monoamine transporter (VMAT2) located in presynaptic vesicles. The purpose of this work was to evaluate various model configurations for analysis of [11C]DTBZ with the aim of providing the optimal measure of monoamine vesicular transporter density obtainable from a single dynamic PET study. PET studies on seven young normal volunteer subjects, ages 20–35, were performed following i.v. injection of 666 ± 37 MBq (18 ± 1 mCi) of (+)-α-[11C]DTBZ. Dynamic acquisition consisted of a 15-frame sequence over 1 h. Analysis methods included both creation of pixel-by-pixel functional images of transport ( K1) and binding ( DVtot) and nonlinear least-squares analysis of volume-of-interest data. Pixel-by-pixel calculations were performed for both two-compartment weighted integral calculations and slope-intercept estimations from Logan plots. Nonlinear least-squares analysis was performed applying model configurations with both two-compartments, estimating K1 and DVtot, and three compartments, estimating K1- k4. For the more complex configuration, we examined the stability of various binding-related parameters including k3 ( kon Bmax′), k3/ k4 ( Bmax′/ Kd), DVsp [( K1/ k2)( k3/ k4)], and DVtot [ K1/ k2(1 + k3/ k4)]. The three-compartment model provided significantly improved goodness-of-fit compared to the two-compartment model, yet did not increase the uncertainty in the estimate of the DVtot. Without constraining parameters in the three-compartment model fits, DVtot was found to provide a more stable estimate of binding density than either k3, k3/ k4, or DVsp. The two-compartment least-squares analysis yielded approximately 10% underestimations of the total distribution. However, this bias was found to be very consistent from region to region as well as across subjects as indicated by the correlation between two- and three-compartment DVtot estimates of 0.997. We conclude that (+)-α-[11C]DTBZ and PET can provide excellent measures of VMAT2 density in the human brain.
Decreased Heterogeneity of Capillary Plasma Flow in the Rat Whisker-Barrel Cortex during Functional Hyperemia
Authors: Vogel Johannes , Kuschinsky Wolfgang
Published: November 2003

The pattern of capillary plasma perfusion was investigated in the rat brain during functional activation. Functional hyperemia was induced in the left whisker-barrel cortex by deflection of the right mystacial vibrissae for 2 min at frequencies of 1–7 Hz. Rats were decapitated under anesthesia 3 s after i.v. bolus injection of Evans blue dye. The steep increase of the arterial dye concentration ensures that divergent capillary plasma transit times result in unequal intracapillary dye concentrations. Plasma perfusion heterogeneity was determined from the coefficient of variation (CV) of Evans blue concentrations measured in numerous single capillaries of the whisker-barrel cortex. Functional hyperemia was quantified from measurements of CBF using the [14C]-iodoantipyrine technique in a second experimental group. CBF in the left whisker-barrel cortex increased with the stimulation frequency and was maximal at 5 Hz compared to the right side. Conversely, plasma perfusion heterogeneity decreased with stimulation frequency in a reciprocal way, being minimal at 5 Hz. Results indicate a decrease in the microcirculatory flow heterogeneity during functional hyperemia in the brain.
Lactate Accumulation during Moderate Hypoxic Hypoxia in Neocortical Rat Brain
Authors: Payen Jean-François , LeBars Emmanuelle , Wuyam Bernard , Tropini Bernard , Pépin Jean-Louis , Lévy Patrick , Décorps Michel
Published: November 2003

Neocortical metabolism was studied during moderate hypoxic hypoxia, reoxygenation, and postmortem periods in anesthetized normocapnic rats using 1H nuclear magnetic resonance (NMR) spectroscopic imaging. Rats were prepared with unilateral common carotid occlusion to determine the ipsilateral metabolic effects of inadequate cerebral blood flow (CBF) response to hypoxia. No difference in brain metabolism between the two hemispheres was found during the control period. Hypoxic hypoxia (Pao2 = 54.1 ± 5.8 mm Hg) resulted in a significant rise in neocortical lactate peak in both hemispheres, with an additional marked rise in the clamped side compared to the unclamped side (53 ± 27 vs. 22 ± 13% of postmortem value, p < 0.001). These lactate changes were not reversible within 30 min of reoxygenation in the clamped hemisphere. No changes in neocortical lactate peak were observed while elevating arterial lactate via intravenous lactate infusion without hypoxia. In addition, hypoxic hypoxia resulted in an apparent decrease in neocortical water and N-acetyl aspartate (NAA) signals, which were related to a shortening in T2 relaxation times. It is concluded that neocortical lactate is an early metabolic indicator during moderate hypoxic hypoxia in normocapnic conditions.
Editing of GluR2 RNA in the Gerbil Hippocampus after Global Cerebral Ischemia
Authors: Rump Andreas , Sommer Clemens , Gass Peter , Bele Sylvia , Meissner Dörte , Kiessling Marika
Published: November 2003

Postischemic delayed neuronal death (DND) in CA1 of the gerbil hippocampus is thought to be caused by an abnormal increase of Ca2+ influx into the cell, mediated by excessive activation of glutamate receptors. One subtype of glutamate receptors, the AMPA receptor, is not permeable to calcium ions as long as an edited form of its GluR2 subunit is present. It is possible that global ischemia interferes with the posttranscriptional editing of the GluR2 mRNA and thus leads to calcium influx via the AMPA receptor. In order to test this hypothesis, we examined the extent of GluR2 RNA editing in CA1 and CA3 microdissected from gerbil hippocampus after 5 min of global ischemia and various recirculation intervals. At each interval tested, quality and quantity of mRNA editing in the vulnerable CA1 region were the same as in CA3. Furthermore, postischemic mRNA editing in both hippocampal regions was indistinguishable from editing in untreated control animals. Our results clearly demonstrate that global ischemia does not cause impairment of GluR2 RNA editing, which is thus not responsible for the abnormal calcium permeability of the postischemic cell membrane.
Cyclo-oxygenase-2 Messenger RNA Induction in Focal Cerebral Ischemia
Authors: Colląo-Moraes Yolanda , Aspey Benjamin , Harrison Michael , de Belleroche Jacqueline
Published: November 2003

We have characterized the induction of the mitogen-inducible form of cyclo-oxygenase, COX-2, during focal cerebral ischemia following permanent middle cerebral artery occlusion (MCAO) in the rat. Marked unilateral induction of COX-2 mRNA was detected in ischemic regions ipsilateral to the occlusion. A significant increase in COX-2 mRNA was detected in “core” and “penumbra” regions of the cerebral cortex between 4 and 24 h after occlusion; this was most marked at 4 h in the penumbra region, in which a 19-fold increase above untreated control levels was detected. A smaller but significant induction was also detected at 4 h in the caudate. A correlation was demonstrated between the extent of COX-2 mRNA induction in cortical regions at 4 h and the severity of tissue damage subsequently detected at 24 h post MCAO. MK-801 significantly attenuated the induction of COX-2 mRNA in the penumbra region at 4 h. The demonstration of COX-2 induction following experimental ischemia highlights the importance of this reaction and its products and by-products, for example, free radicals, in the tissue response to this insult.
Reduction in Posterior Cerebral Artery Blood Flow Velocity during Caloric Vestibular Stimulation
Authors: Tiecks F. P. , Planck J. , Haberl R. L. , Brandt T.
Published: November 2003

Transcranial Doppler (TCD) sonography was performed to assess the differential effects of caloric vestibular stimulation on cerebral blood flow (CBF) to the vestibular [middle cerebral artery (MCA)] and the visual [posterior cerebral artery (PCA)] cortices in 15 healthy volunteers with eyes closed. Caloric irrigation caused a sensation of rotation and nystagmus and not only increased velocity in the MCA (2.7 ± 3.3%) but also, simultaneously, decreased velocity in the PCA (4.2 ± 6.2%, p < 0.001 vs. MCA). CBF changes indicated by changes in velocity through the MCA and PCA may reflect functional deactivation of the visual cortex together with activation of the vestibular cortex: this beneficial multisensory interaction suppresses distressing oscillopsia.
Effects of the AMPA-Receptor Antagonist, NBQX, on Neuron Loss in Dentate Hilus of the Hippocampal Formation after 8, 10, or 12 Min of Cerebral Ischemia in the Rat
Authors: Hu Ping , Diemer Nils Henrik , Bruhn Torben , Johansen Flemming Fryd
Published: November 2003

The α-amino-3-hydroxy-5-methyl-4-isoxazole (AMPA) receptor antagonist, 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo( F)quinoxaline (NBQX), offers protection to hippocampal CA1 pyramidal cells after short episodes of transient cerebral ischemia. Besides CA1 pyramidal cells, neurons containing somatostatin (SS) and located in the dentate hilus of the hippocampal formation are lost after cerebral ischemia. We studied the protective effects of NBQX on SS neurons in the hilus and on hippocampal CA1 pyramidal cells following 8, 10, or 12 min of four-vessel occlusion ischemia during systemic hypotension. NBQX was administered 3 × 30 mg/kg at 0, 10, and 25 after induction of ischemia or sham, and all rats survived for 7 days. NBQX given to control rats without ischemia had no influence on number or morphology of hilar SS neurons and CA1 pyramidal cells. After 8 min of ischemia, NBQX prevented loss of hilar SS neurons. After 10 and 12 min of ischemia, NBQX had no significant effects on loss of SS neurons in the dentate hilus. However, in all ischemic groups, NBQX significantly reduced loss of CA1 pyramidal cells as compared to control rats. This neuroprotective effect decreased gradually and significantly as the time of ischemia increased. Our results support the observation that SS neurons in hilus are among the most ischemia-vulnerable neurons in the brain. We found that administration of NBQX in generally accepted dosages can protect the rapidly dying SS neurons in hilus from only brief episodes of ischemia.
Isoflurane and Propofol Block Neurotoxicity Caused by MK-801 in the Rat Posterior Cingulate/Retrosplenial Cortex
Authors: Jevtović-Todorović Vesna , Kirby Charity O. , Olney John W.
Published: November 2003

In acute brain injury syndromes, the potent N-methyl-D-aspartate (NMDA) antagonist, MK-801, can prevent neuronal degeneration, and the general anesthetics, isoflurane and propofol, may also provide neuroprotective benefits. An obstacle to the use of NMDA antagonists for neuroprotective purposes is that they can cause a neurotoxic vacuole reaction in cerebrocortical neurons. This study demonstrates the ability of isoflurane and propofol to prevent the neurotoxic vacuole reaction induced by MK-801. Low sedative doses of inhaled isoflurane (1%) or intravenous (i.v.) propofol (7.5 mg/kg/h) were as effective as higher general anesthetic doses. Thus, in the clinical management of acute brain injury conditions such as stroke and brain trauma, administration of one of these anesthetic agents together with an NMDA antagonist may be an excellent formula for obtaining optimal neuroprotection while eliminating serious side effects.
Rapid Preconditioning Protects Rats against Ischemic Neuronal Damage after 3 but Not 7 Days of Reperfusion following Global Cerebral Ischemia
Authors: Pérez-Pinzón Miguel A. , Xu Guang-Ping , Dietrich W. Dalton , Rosenthal Myron , Sick Thomas J.
Published: November 2003

Earlier studies indicated that sublethal ischemic insults separated by many hours may “precondition” and, thereby, protect tissues from subsequent insults. In Wistar rats, we examined the hypothesis that ischemic preconditioning (IPC) can improve histopathological outcome even if the “conditioning” and “test” ischemic insults are separated by only 30 min. Normothermic (36.5–37°C) global cerebral ischemia was produced by bilateral carotid artery ligation after lowering mean systemic blood pressure. The conditioning ischemic insult lasted 2 min and was associated with a time sufficient to provoke “anoxic depolarization” (AD) (i.e., the abrupt maximal increase in extracellular potassium ion activity). After 30 min of reperfusion, 10-min test ischemia was produced, and histopathology was assessed 3 and 7 days later. After 3 days of reperfusion, neuroprotection was most robust in the left lateral, middle and medial subsections of the hippocampal CA1 subfield and in the cortex, where protection was 91, 76, 70 and 86%, respectively. IPC also protected the right lateral, middle and medial subsections of the hippocampal CA1 region. These data demonstrate that neuroprotection against acute neuronal injury can be achieved by conditioning insults followed by only short (30 min) periods of reperfusion. However, neuroprotection almost disappeared when reperfusion was continued for 7 days. When test ischemia was decreased to 7 min, a clear trend of neuroprotection by IPC was observed. These data suggest that subsequent rescue of neuronal populations could be achieved with better understanding of the neuroprotective mechanisms involved in this rapid IPC model.
Identification, Characterization, and Functional Role of Phosphodiesterase Type IV in Cerebral Vessels: Effects of Selective Phosphodiesterase Inhibitors
Authors: Willette Robert N. , Shiloh Aaron O. , Sauermelch Charles F. , Sulpizio Anthony , Michell Marcus P. , Cieslinski Lenora B. , Torphy Theodore J. , Ohlstein E. H.
Published: November 2003

The role of the phosphodiesterase type IV isozyme (PDE IV) in the regulation of cerebrovascular tone was investigated in the canine basilar artery in vitro and in vivo. The PDE isozymes extracted from the canine basilar artery were isolated by diethylaminoethanol (DEAE)-Sepharose affinity chromatography and identified based on sensitivity to tsozyme-selective PDE inhibitors. [3H]cAMP hydrolysis was observed in one major and one minor peak of activity. The predominant peak was inhibited by the addition of cGMP (25%), siguazodan (26%), rolipram (39%), and the combination of siguazodan and rolipram (95%). Selective PDE IV inhibitors BRL 61063, rolipram, and denbufylline were equieffective inhibitors of [3H]-ccAMP hydrolysis mediated by PDE IV isolated from the canine basilar artery [concentrations producing 50% inhibition (IC50s) = 0.21 ± 0.05 μM, 0.67 ± 0.23 μM, and 0.73 ±0.16 μM, respectively]. In precontracted isolated ring segments of the canine basilar artery, selective PDE IV inhibitors produced potent and complete relaxation (IC50s <150 nM). In contrast, zaprinast (a selective PDE V inhibitor) and siguazodan (a selective PDE III inhibitor) produced only weak relaxation of the basilar artery (IC50s = 4.5 μM and >10 μM, respectively). Vasorelaxation produced by PDE IV inhibitors was not altered by removing the endothelium, l-NAME, or adenosine receptor antagonism. In a canine model of acute cerebral vasospasm, all three selective PDE IV inhibitors reversed basilar artery spasm produced by autologous blood without altering mean arterial blood pressure. In contrast, prolonged treatment with BRL 61063 failed to alter the development of basilar spasm in the two hemorrhage canine models of chronic cerebral vasospasm. Denbufylline-induced relaxation in vitro was also significantly impaired in basilar arteries obtained from the model of chronic vasospasm. In conclusion, PDE IV appears to be the predominant isozyme regulating vascular tone mediated by cAMP hydrolysis in cerebral vessels. In addition, vasorelaxation modulated by PDE IV is compromised in chronic cerebral vasospasm associated with subarachnoid hemorrhage.