Transient cerebral ischemia. Association of apoptosis induction with hypoperfusion.

Apoptosis is thought to be important in the pathogenesis of cerebral ischemia. The mechanism of apoptosis induction remains unclear but several studies suggest that it is preferentially triggered by mild/moderate microcirculatory disturbances. We examined in cats whether induction of apoptosis after 2.5 h of unilateral middle cerebral artery occlusion plus 10 h of reperfusion is influenced by the degree of cerebral microcirculatory disturbance. Quantitative monitoring over time of the disturbances of cerebral microcirculation in ischemic brain areas and evaluation of cytotoxic edema associated with perfusion deficits was achieved by using two noninvasive magnetic resonance imaging techniques: (a) high-speed echo planar imaging combined with a bolus of magnetic susceptibility contrast agent; and (b) diffusion-weighted imaging. Apoptosis-positive cells were counted in anatomic areas with different severity of ischemic injury characterized by magnetic resonance imaging, triphenyltetrazolium chloride, and hemotoxylin and eosin staining. The number of apoptosis-positive cells was significantly higher in anatomic areas with severe perfusion deficits during occlusion and detectable histologic changes 10 h after reperfusion. In contrast, in areas where perfusion was reduced but maintained during occlusion there were no detectable histological changes and significantly fewer apoptosis-positive cells. A similar number of cells that undergo apoptosis were shown in regions with transient or prolonged subtotal perfusion deficits. These results suggest that the apoptotic process is induced in the ischemic core and contributes significantly in the degeneration of neurons associated with transient ischemia.

Measurement of ventricular volumes in the dog by nuclear magnetic resonance imaging.

Nuclear magnetic resonance (NMR) imaging is a new, noninvasive approach for imaging the cardiovascular system. Being a three-dimensional technique, NMR imaging has the capability of measuring volumes without the need for assumptions regarding ventricular geometry. In this study, the technique was validated in 19 excised dog hearts, filled with silicone-rubber, imaged using a multislice spin-echo sequence. The volume of the cavity in each slice was calculated from the number of pixels outlined for each slice multiplied by the pixel volume. Ventricular volumes measured by NMR imaging were highly correlated with cast volumes measured by water displacement: right ventricle (RV):RVNMR = 1.05 RVcast - 1.62; r = 0.99, SEE = 0.96 ml; left ventricle (LV):LVNMR = 0.98 LVcast + 0.35, r = 0.98, SEE = 1.48 ml. After validation in casts, NMR imaging volumes were measured in eight living dogs using a multiphasic gated technique to obtain images at 5, 105, 205, 305 and 405 ms after the QRS complex. Cardiac output (CO) and stroke volume (SV) measured by NMR imaging were significantly correlated with thermodilution (TD) measurements (CONMR = 0.63 COTD + 0.51 liters/min; r = 0.78, SEE = 0.57 liters/min; SVNMR = 0.67 SVTD + 1.95 ml; SEE = 5.58 ml). Right and left stroke volumes were closely related (LVSVNMR = 0.9 RVSVNMR + 1.75; r = 0.94, SEE = 4.32 ml), with the slope and intercept of the regression line showing no difference from 1 and 0, respectively. However, volumes determined by NMR imaging underestimated the thermodilution measurements, presumably reflecting the inability to obtain a true systolic image with the present sampling rate.(ABSTRACT TRUNCATED AT 250 WORDS)

Influence of severity of myocardial injury on distribution of macromolecules: extravascular versus intravascular gadolinium-based magnetic resonance contrast agents.

OBJECTIVES: This study sought to 1) compare the distribution of extravascular (573 Da) and intravascular (92 kDa) magnetic resonance (MR) contrast agents in reperfused infarcted myocardium, and 2) investigate the effect of injury severity on these distribution patterns. BACKGROUND: Myocardial distribution of low and high molecular weight contrast agents depends on vascular permeability, diffusive/convective transport within the interstitium and accessibility of the intracellular compartment (cellular integrity). METHODS: To vary the severity of myocardial injury, 72 rats were subjected to 20, 30, 45 or 75 min (n = 18, respectively) of coronary artery occlusion. After 2 h of reflow, the animals received either 0.05 mmol/kg of gadolinium-diethylenetriaminepentaacetic acid-bismethylamide (Gd-DTPA-BMA) (n = 24), (Gd-DTPA)30-albumin (n = 24) or saline (control group, n = 24). Three minutes after injection, the hearts were excised and imaged (spin-echo imaging parameters: repetition time 300 ms, echo time 8 ms, 2-tesla system), followed by triphenyltetrazolium chloride staining for infarct detection and sizing. RESULTS: Histomorphometric and MR infarct size (expressed as percent of slice surface) correlated well: r = 0.96 for Gd-DTPA-BMA; r = 0.95 for (Gd-DTPA)30-albumin. On Gd-DTPA-BMA-enhanced images, reperfused myocardial infarctions were homogeneously enhanced. The ratio of signal intensity of infarcted/ normal myocardium increased with increasing duration of ischemia (overall p < 0.0001, analysis of variance [ANOVA]), indicating an increase in the distribution volume of Gd-DTPA-BMA in postischemic myocardium. On (Gd-DTPA)30-albumin-enhanced images, reperfused infarctions consisted of a bright border zone and a less enhanced central core. The extent of the core increased with increasing duration of ischemia (overall p value < 0.0001, ANOVA). CONCLUSIONS: At 2 h of reperfusion, the distribution of MR contrast agents in postischemic myocardium is 1) specific for extravascular and intravascular agents, and 2) modulated by the duration of ischemia.

Effects of nicardipine, a calcium antagonist, on myocardial salvage and high energy phosphate stores in reperfused myocardial injury.

The current study determined the effectiveness of nicardipine, a 1,4-dihydropyridine calcium antagonist, in preserving reperfused myocardium in a cat model of temporary coronary occlusion and ascertained if replenishment of myocardial phosphate stores during reperfusion as defined by phosphorus-31 nuclear magnetic resonance (NMR) spectroscopy was indicative of salvage. Twenty open chest, anesthetized cats were studied with use of a snare ligature around the proximal left anterior descending coronary artery, with a coil sutured to the epicardial surface overlying the distribution of the artery. Peak areas of phosphocreatine, inorganic phosphate and adenosine triphosphate (ATP) NMR signals were measured during 1 h of occlusion followed by 1.5 h of reperfusion. Infarct size and jeopardy area were determined in vitro by simultaneous infusion of phthalocyanine blue dye and triphenyltetrazolium chloride into the aorta and the left anterior descending coronary artery, respectively, after 5 h of myocardial reperfusion. Nicardipine-treated and control groups had similar jeopardy area values (41.2 +/- 1.6% versus 47.4 +/- 3.1% of the left ventricle), but infarct area was significantly reduced in the nicardipine-treated group (3.2 +/- 1.1% versus 24.9 +/- 7.5% of jeopardy area, p less than 0.01). High energy phosphate compounds remained markedly altered during reperfusion in both groups. No significant improvement in phosphocreatine or inorganic phosphate recovery was observed in animals pretreated with nicardipine despite an 87% reduction in infarct size. Myocardial ATP was greater during reperfusion in the nicardipine-treated compared with the control group (average over initial 90 min of reperfusion 58 +/- 6% versus 46 +/- 3% of baseline values, p less than 0.05), suggesting improved recovery of ATP. However, the measured levels of high energy phosphate compounds during reperfusion and their ratios did not correlate with infarct size and thus were not predictive of myocardial salvage.

31P magnetic resonance spectroscopy of pressure overload hypertrophy in rats: effect of reduced perfusion pressure.

STUDY OBJECTIVE - The purpose of the study was to confirm the presence of abnormalities in the coronary vessels of hypertensive hearts, and to examine the effects of reduced coronary perfusion pressure. DESIGN - Rats were made hypertensive by aortic banding, after which coronary flow and myocardial energy metabolites were studied in isolated hearts at physiological (140 cm H2O) and reduced (80 cm H2O) coronary perfusion pressures and compared with normotensive controls. SUBJECTS - Wistar-Kyoto rats between 250 and 300 g were used. Left ventricular hypertrophy was generated by aortic banding in 29 rats; 8 were studied one week after banding, and 21 three weeks after banding. There were 45 controls. MEASUREMENTS and RESULTS - Energy metabolites were assessed using 31P magnetic resonance spectroscopy, standardised by high performance liquid chromatography of rapidly freeze clamped tissue. Left ventricular wall thickness was determined using two dimensional echocardiography. Coronary flow (normalised for heart weight) was reduced significantly after one and three weeks of left ventricular hypertrophy, and at either physiological or below physiological pressures. Hearts from aortic banded animals developed higher intraventricular pressure with reduced oxygen consumption when perfused at a physiological pressure, indicating increased thermodynamic efficiency. When perfused at reduced pressure, the developed pressure declined significantly in both the one week and the three week banded groups compared to normal hearts. The phosphorylation potential and intracellular pH (pHi) were not significantly lower after one week and three weeks of left ventricular hypertrophy when perfused at physiological pressure. When perfused at reduced pressure, phosphorylation potential declined significantly in both groups of hypertrophied hearts, whereas pHi declined significantly only in the three week hypertrophy group. CONCLUSIONS - There is improved thermodynamic efficiency of the hypertrophied myocardium when perfused at a physiological pressure, but when perfused at a reduced pressure, ventricular function, phosphorylation potential and pHi decline in rat hearts after three weeks of aortic constriction, indicating an impairment of coronary reserve.

Reperfusion differentially induces caspase-3 activation in ischemic core and penumbra after stroke in immature brain.

BACKGROUND AND PURPOSE: Different strategies for neuroprotection of neonatal stroke may be required because the developing brain responds differently to hypoxia-ischemia than the mature brain. This study was designed to determine the role of caspase-dependent injury in the pathophysiology of pure focal cerebral ischemia in the immature brain. METHODS: Postnatal day 7 rats were subjected to permanent or transient middle cerebral artery (MCA) occlusion. Diffusion-weighted MRI was used during occlusion to noninvasively map the evolving ischemic core. The time course of caspase-3 activation in ischemic brain tissue was determined with the use of an Asp-Glu-Val-Asp-aminomethylcoumarin cleavage assay. The anatomy of caspase-3 activation in the ischemic core and penumbra was mapped immunohistochemically with an anti-activated caspase-3 antibody in coronal sections that matched the imaging planes on diffusion-weighted MRI. RESULTS: A marked increase in caspase-3 activity occurred within 24 hours of reperfusion after transient MCA occlusion. In contrast, caspase-3 activity remained significantly lower within 24 hours of permanent MCA occlusion. Cells with activated caspase-3 were prominent in the penumbra beginning at 3 hours after reperfusion, while a more delayed but marked caspase-3 activation was observed in the ischemic core by 24 hours after reperfusion. CONCLUSIONS: In the neonate, caspase-3 activation is likely to contribute substantially to cell death not only in the penumbra but also in the core after ischemia with reperfusion. Furthermore, persistent perfusion deficits result in less caspase-3 activation and appear to favor caspase-independent injury.

High-speed MR imaging of ischemic brain injury following stenosis of the middle cerebral artery.

Magnetic susceptibility contrast-enhanced and diffusion-weighted echo planar magnetic resonance (MR) imaging was performed using a cat model of acute regional cerebral ischemia induced by partial stenosis of the right middle cerebral artery (MCA). The imaging data were correlated with triphenyltetrazolium chloride (TTC)-stained histopathologic coronal brain sections to determine the prognostic efficacy of high-speed MR imaging techniques in differentiating mild, moderate, and severe cerebral hypoperfusion. Brains of animals without cortical injury on TTC staining were found to have a reduction in peak contrast enhancement of 32 +/- 6% (mean +/- SD) below control values with no significant change in the apparent diffusion coefficient (ADC), determined from the diffusion-weighted MR images. In cases where moderate ischemic injury was observed in the TTC-stained sections, a 10-20% drop in the ADC was found over the 6-h study period, accompanied by a much wider variation in peak contrast enhancement. Finally, where TTC staining showed severe ischemic brain damage, a 40-50% drop in ADC and a reduction in peak contrast enhancement effect of > 95% were observed as early as 1 h following MCA stenosis. The significant correlation between imaging observations and histologically confirmed cerebral ischemia indicates that magnetic susceptibility contrast-enhanced echo planar MR imaging is sensitive to slight reductions in cerebral perfusion that fall below the threshold for reliably detectable ischemia-induced alterations in ADC. First-pass perfusion-sensitive imaging may thus be diagnostically useful in differentiating severely hypoperfused permanently injured tissue from the mildly hypoperfused ischemic penumbra.

Effect of lidocaine on acute regional myocardial ischemia and reperfusion in the cat. An in-vivo 31P magnetic resonance spectroscopy study.

RATIONALE AND OBJECTIVES: The authors examined the relationship between myocardial infarction, high-energy phosphate compounds, and regional contractility after myocardial ischemia and reperfusion in cats. METHODS: Hemodynamic measurements, high-energy phosphate levels, and segmental shortening were measured every 30 minutes in two groups of cats subjected to 2 hours of occlusion of the left anterior descending coronary artery and 4 hours reperfusion. Group 1 (n = 10) animals were infused with a low level of lidocaine, 0.05 mg/kg/hr, while group 2 (n = 10) received a higher dose, 7.5 mg/kg/hr. The infarcted region was measured postmortem. RESULTS: Group 1 animals had larger infarcts (39 +/- 6 vs. 12 +/- 5% of jeopardy, P < .05) and less phosphocreatine recovery during reflow (52 +/- 7% vs. 73 +/- 2% of control, P < .01) than did group 2. Group 2 showed recovery of percentage systolic shortening during reflow (1.4 +/- 2% at 30 minutes vs. 7.1 +/- 2.3% at 4 hours, P < .05), whereas group 1 exhibited no improvement. A significant correlation was found between infarct size under the surface coil and phosphocreatine content during reflow, but not between contractile function and infarction size or metabolite levels during reflow. CONCLUSIONS: Lidocaine infusion enhanced recovery of myocardial contractility during reperfusion and decreased infarct size. Greater recovery of phosphocreatine during reperfusion was predictive of greater myocardial salvage during reperfusion.

The use of contrast-enhanced magnetic resonance imaging to define ischemic injury after reperfusion. Comparison in normal and hypertrophied hearts.

RATIONALE AND OBJECTIVES: Magnetic resonance imaging (MRI) was used to demonstrate the infarction size in reperfused ischemic myocardium of normal and hypertrophied hearts, and to test the hypothesis that hypertrophied hearts manifest greater susceptibility to ischemia. METHODS: Normal rats (n = 11) and rats subjected to left ventricular hypertrophy (LVH) by aortic banding (n = 13) were studied. After 7 weeks, the left coronary artery was occluded for 25 minutes and reperfused for 1 hour before MRI. Electrocardiogram-gated spin-echo images were acquired before and after administration of 0.3 mmol/kg gadoteridol. To quantify the hyperintense area demarcated by gadoteridol, 3 transaxial images were acquired at different levels. Jeopardy and infarcted areas were measured in the same three slices postmortem using blue dye and triphenyltetrazolium chloride (TTC) stain, respectively. RESULTS: Before administration, there was no significant difference in signal intensity between nonischemic (0.42 +/- 0.03 arbitrary units) and ischemic (0.41 +/- 0.03) myocardium in either group. After gadoteridol injection, signal intensity of the reperfused injured region was higher than that of nonischemic myocardium (1.48 +/- 0.16 vs. 0.72 +/- 0.06, P < .05). Magnetic resonance delineation of the hyperintense area persisted for at least 30 minutes. The size of the hyperintense area was larger in LVH than in control hearts (25 +/- 5% vs. 7 +/- 3% of LV surface area, P < .05) and did relate closely to the area of myocardial infarction (r = .97), but not with the jeopardy area (r = .42). On TTC staining, the infarction size also was significantly greater in LVH than in normal group (18 +/- 5% vs. 5 +/- 2% of LV surface area, P < .05). The jeopardy areas of normal and LVH hearts showed no significant difference (46 +/- 2% vs. 47 +/- 3%). CONCLUSION: Magnetic resonance imaging confirms the concept that reperfused myocardial injury is larger in LVH than normal hearts after brief coronary occlusion. Contrast-enhanced MRI can define the size of reperfused myocardial injury. Thus, MRI is a suitable technique to assess conditions accentuating ischemic injury.

Real-time dynamics of an extravascular magnetic resonance contrast medium in acutely infarcted myocardium using inversion recovery and gradient-recalled echo-planar imaging.

RATIONALE AND OBJECTIVES: The purposes of this study are to evaluate the first-pass profile of gadolinium-BOPTA/Dimeg (Gd-BOPTA/Dimeg) during its transit through hearts subjected to acute myocardial infarction, and to delineate these infarcted regions by the use of ultrafast magnetic resonance imaging (MRI). METHODS: Regional ischemia was induced in anesthetized rats by occluding the left coronary artery. Imaging parameters for single shot EPI included TE, 10 mseconds; AT, 33 mseconds; and 64 x 64-pixel matrix. Consecutive images were obtained every 1 to 2 seconds over a 30-second period. After approximately two images, Gd-BOPTA/Dimeg was injected intravenously (0.05 and 0.25 mmol/kg). RESULTS: Gd-BOPTA/Dimeg (0.05 mmol/kg), with inversion recovery EPI, produced a substantial increase in signal intensity of right and then left ventricular blood. Normally perfused myocardium also was enhanced, but not the acutely infarcted region. Clear delineation of the infarcted region as negatively enhanced "cold spots" persisted for at least 20 seconds. Gd-BOPTA/Dimeg (0.25 mmol/kg) with standard gradient-recalled EPI produced a different profile of signal intensity changes. Signal intensities of ventricular blood and normal myocardium were greatly reduced, leaving the infarcted zone as a positively enhanced "hot spot." Delineation of the infarcted region persisted for 6 to 8 seconds. The infarcted zone detected with MRI corresponded to that observed at autopsy. CONCLUSIONS: Regions of acute myocardial infarction can be detected as negatively enhanced "cold spots" or positively enhanced "hot spots" by studying the first-pass dynamics of Gd-BOPTA/Dimeg through hearts with regional ischemia by use of single shot EPI.

Preservation of high-energy phosphate reserves in a cat model of post-ischemic myocardial dysfunction.

Brief episodes of myocardial ischemia are known to cause reversible depression of regional myocardial contraction after reperfusion. One of the mechanisms of this persistent regional dysfunction has been proposed to be depletion of high-energy phosphate compounds. Eight cats were prepared with a reversible snare occluder around the left anterior descending artery (LAD); a surface coil sutured to the epicardial surface over the LAD territory for measurement of 31-phosphorus (31P) magnetic resonance spectroscopy (MRS) spectra; and a pair of ultrasonic crystals implanted in the mid-myocardium for measurement of regional segment length shortening. The baseline value of percent segment length shortening (%SS) was 12.8 +/- 1.4%. Increased afterload did not significantly alter high-energy phosphate levels or %SS. All animals exhibited passive systolic bulging during occlusion (-8.4 +/- 3.6% systolic shortening) as well as reduced phosphocreatine (PRc, 30 +/- 3% of control) and increased inorganic phosphorus (Pi) (239 +/- 18%), but there was no change in adenosine triphosphate (ATP). During reflow, %SS did not completely recover (4.0 +/- 2.9%, P less than .05 versus baseline). PCr and Pi returned to control levels during the first 30 minutes of reperfusion. Increased afterload had no significant effect on high-energy phosphates or %SS in stunned hearts. These findings indicate a lack of correlation between recovery of high-energy phosphate stores and regional myocardial contractility in stunned myocardium. High-energy phosphate reserves are preserved in stunned myocardium and are unlikely to be a direct cause of myocardial dysfunction.

Magnetic resonance imaging of acute myocardial ischemia using a manganese chelate, Mn-DPDP.

Nine adult rats underwent occlusion of the left coronary artery (LCA) to assess the ability of a manganese chelate of N, N'-Bis (pyridoxal-5-phosphate) ethylenediamine-N, N'-diacetic acid (Mn-DPDP), to delineate acute myocardial ischemia. Hemodynamic effects of the contrast medium were tested in the isolated rat heart. Gated transaxial images of the heart at the mid-ventricular level were obtained using a 2 Tesla magnet. A TE of 20 msec and a TR of 1 R-R interval (approximately 250 msec) were used. Images were taken prior to injection of 400 mumol/kg of Mn-DPDP, 2 minutes after injection, and then at 15 minute intervals for one hour. The time between LCA occlusion and injection of contrast averaged 95 minutes. The signal intensity (SI) of normal myocardium was increased by 125 +/- 9% immediately after injection, and did not significantly vary over one hour. SI of ischemic myocardium increased by only 16 +/- 14% immediately after injection, gradually rising to 44 +/- 13% after one hour. Visual discrimination between normal and ischemic myocardium was obtained throughout the study. The percent contrast between normal and ischemic myocardium was 47.2 +/- 6% at 2 minutes after injection, gradually decreasing to a final value of 31.3 +/- 4%. No hemodynamic effects were produced in the isolated heart using concentrations in the perfusate several-fold higher than those expected to be produced by the intravenous injection of 400 mumol/kg Mn-DPDP. Mn-DPDP shows the potential for delineation of the jeopardy area resulting from coronary occlusion for at least one hour after injection of the agent.

Comparison of T1-enhancing and magnetic susceptibility magnetic resonance contrast agents for demarcation of the jeopardy area in experimental myocardial infarction.

RATIONALE AND OBJECTIVES: This study compared the areas demarcated by a T1-enhancing agent, Gd-DTPA-BMA, and a magnetic susceptibility agent, Dy-DTPA-BMA, with 201thallium autoradiography (indicator of perfusion) and postmortem histochemical staining with triphenyltetrazolium chloride (TTC)(indicator of infarction). METHODS: Thirteen rats were subjected to coronary artery occlusion for 3 to 4 hours before acquisition of four sets of electrocardiogram-gated spin-echo magnetic resonance (MR) images: T1-weighted images before and after 0.2 mmol/kg Gd-DTPA-BMA; and T2-weighted images before and after 0.3 mmol/kg Dy-DTPA-BMA. After MR imaging, intravenous 201thallium delineated the area of decreased myocardial perfusion. At autopsy, TTC staining delineated the area of myocardial infarction. RESULTS: A myocardial region in the distribution of the occluded artery was delinated as a hyperintense area ("hot-spot") by Dy-DTPA-BMA and as a hypointense area ("cold-spot") by Gd-DTPA-BMA. The hyperintense area demarcated by Dy-DTPA-BMA (51 +/- 3% of the area of the midequitorial slice of the left ventricle) showed a closer relationship to the area of decreased myocardial perfusion (jeopardized area) (46 +/- 3%), determined by 201thallium autoradiography, than the area of myocardial infarction (36 +/- 4%), determined by histochemical staining. However, the hypointense area demarcated by Gd-DTPA-BMA (29 +/- 2%) did not relate as closely to the area of decreased myocardial perfusion (slope = 0.54) or the area of myocardial infarction (r = 0.46). CONCLUSIONS: The abnormal myocardial area delineated by the magnetic susceptibility agent showed a closer relationship to the area of deficient myocardial perfusion (jeopardy area) after coronary occlusion than that defined by T1-enhancing contrast media.

Neonatal reversible focal cerebral ischemia: a new model.

While numerous animal models exist for studying neonatal brain injury after cerebral ischemia-hypoxia, an adequate model for assessing reversible focal ischemia in the neonatal rat has not been reported. This paper describes in detail a new surgical procedure for creating a non-hemorrhagic, reperfused focal ischemic lesion in the neonatal, 7-day-old rat pup.

Mild hypothermia decreases the incidence of transient ADC reduction detected with diffusion MRI and expression of c-fos and hsp70 mRNA during acute focal ischemia in rats.

The effects of mild hypothermia on the apparent diffusion coefficient of water (ADC) and expression of c-fos and hsp70 mRNA were examined during acute focal cerebral ischemia. Young adult rats were subjected to 60-min middle cerebral artery occlusion under either normothermia (37.5 degrees C) or hypothermia (33 degrees C). Diffusion-weighted echo-planar magnetic resonance imaging was used to monitor changes in ADC throughout the ischemic period. Perfusion MRI with dysprosium contrast was used at the end of the ischemic period to verify that the occlusion was successful. C-fos and hsp70 mRNA expression were examined with in situ hybridization at the end of the ischemic period. The results indicate that the size of the region that exhibited reduced ADC was smaller during hypothermia than during normothermia. Hypothermia also decreased the frequency of occurrence of transient ADC reductions, especially in dorsal aspects of cortex. Expression of both c-fos and hsp70 mRNA were markedly reduced by hypothermia. Transient ADC reduction and c-fos expression are associated with spreading depression, which is believed to contribute to lesion expansion during acute focal ischemia. The results suggest that part of the neuroprotective effect of hypothermia may be due to a reduced incidence of spreading depression.

Evaluation of recombinant human basic fibroblast growth factor (rhbFGF) as a cerebroprotective agent using high speed MR imaging.

The potential cerebroprotective effects of recombinant human basic fibroblast growth factor (rhbFGF) were evaluated in a feline model of acute cerebral ischemia using high-speed magnetic resonance imaging (MRI) in conjunction with immunohistology. The neuroprotective efficacy of three doses of rhbFGF was evaluated in a unilateral middle cerebral artery (MCA) occlusion/reperfusion model. Ten h following a 2 h period of MCA occlusion in control (vehicle-treated) animals, cerebral perfusion in the ischemic hemisphere was 58 +/- 17% of the contralateral normal hemisphere. Corresponding ischemic/normal perfusion ratios in rhbFGF-treated groups were not significantly different: 54 +/- 16% (14 micrograms/kg/h dose), 40 +/- 19% (42 micrograms/kg/h dose) and 75 +/- 8% (125 micrograms/kg/h dose). Triphenyltetrazolium chloride histopathological assessment demonstrated brain damage in vehicle-treated animals comprising 31 +/- 15% of the hemisphere; in rhbFGF-treated groups injury was not significantly different: 19 +/- 4% (14 micrograms/kg/h rhbFGF), 24 +/- 6% (42 micrograms/kg/h rhbFGF) and 16 +/- 10% (125 micrograms/kg/h rhbFGF). Immunohistochemical analysis of brain sections using glial fibrillary acidic protein (GFAP) revealed that in animals that showed marked perfusion deficits throughout the entire experiment (regardless of treatment), GFAP staining was elevated contralateral to the occlusion and absent ipsilaterally. While some tendency towards protection is found, particularly at higher doses of rhbFGF, it must be concluded that in the chosen stroke model and time interval, the doses used did not significantly improve reperfusion or confer significant cerebroprotective benefit. Non-invasive high-speed MRI was found to be useful for evaluation of putative cerebroprotective agents.

Transient MRI-detected water apparent diffusion coefficient reduction correlates with c-fos mRNA but not hsp70 mRNA induction during focal cerebral ischemia in rats.

Cerebral ischemia induces immediate early genes such as c-fos and stress genes such as hsp70. In this study, the spatial relationships between c-fos and hsp70 mRNA expression and changes detectable with diffusion and perfusion magnetic resonance (MR) imaging were examined. The middle cerebral artery (MCA) of young adult rats was occluded for 30 or 60 min. Diffusion MR (D-MR) images were acquired continuously during the ischemic period and dysprosium-contrast perfusion (P-MR) images were acquired at the end of the ischemic period. C-fos and hsp70 mRNA expression were examined with in situ hybridization. The most significant finding of this work was that for both durations of ischemia, c-fos induction was observed in cortical and sub-cortical regions exhibiting a transient reduction in the apparent diffusion coefficient of water (ADC). Transients which occurred on a time scale of 3 min may have been caused by spreading depression. Those occurring on a 10-min time scale may have been caused by an initial reduction in blood flow with occlusion that was followed by an ischemia-induced increase in collateral blood flow. P-MR imaging showed that perfusion in c-fos positive regions was higher than in regions with persistently reduced ADC. Hsp70 induction did not correlate with transient ADC reduction. It was induced in the MCA territory in regions showing persistent ADC changes, with induction being greatest at the periphery of these regions. It was also induced in regions that exhibited both spontaneous reversal of the diffusion changes and decreased perfusion.

Hyperglycemia augments ischemic brain injury: in vivo MR imaging/spectroscopic study with nicardipine in cats with occluded middle cerebral arteries.

Hyperglycemia is often associated with an increased frequency of cerebrovascular disease and exacerbation of neuronal injury in focal ischemic cerebral infarction. We used a combination of high-field proton MR imaging and 1H and 31P MR spectroscopy to investigate whether hyperglycemia would adversely influence cerebral metabolism and eventual infarct size following unilateral occlusion of the middle cerebral artery (MCA) of cats pretreated with the calcium channel blocker nicardipine. Normoglycemic animals injected with 10 micrograms/kg of nicardipine (8 micrograms.kg-1.hr-1 maintenance dose) manifested only mild disturbances in phosphorus metabolism and cerebral pH regulation compared with untreated controls, and showed a significant reduction in infarct size 7 hr after MCA occlusion. By comparison, hyperglycemic cats (plasma glucose, 200-300 mg/dl) had significantly reduced cerebral high-energy phosphates, elevated lactic acid, and larger ischemic lesions in the occluded MCA territory, irrespective of whether they were treated with nicardipine. These results indicate that moderate hyperglycemia can exaggerate ischemic brain damage by enhancing formation of tissue lactic acid and impairing normal phosphorus metabolism. One implication of this study is that dextrose should not be provided to patients with acute ischemic stroke.

MR imaging of acute experimental ischemia in cats.

The evolution of acute cerebral ischemia was documented by magnetic resonance (MR) imaging in 13 mongrel cats with occlusion of the middle cerebral artery through a transorbital approach. The animals were imaged under anesthesia at intervals from 30 min to 10 days after production of the lesion. An MR imager operating at 0.35 T was used with multislice, multi-spin-echo technique (TR = 500-2000 msec; TE = 28, 56 msec). The animals were sacrificed after imaging for pathologic correlation. Infarcts beyond 4 hr of age were visualized in all subjects. The earliest infarct was seen at 30 min (two cats) as an area of high signal intensity on T2-weighted images. In three other cats, however, 3-hr-old infarcts were not detectable. In one animal, a hemorrhage within a 1-week-old area of infarction was not characterized by MR imaging but was identified on CT scanning. The mass effect of the infarction appeared greatest at 2-4 days after infarction. The basal ganglia showed ischemic effects to best advantage. MR imaging offers previously unavailable sensitivity for the early noninvasive detection of cerebral ischemia in vivo.

Diffusion-weighted MR imaging of acute stroke: correlation with T2-weighted and magnetic susceptibility-enhanced MR imaging in cats.

We evaluated the temporal and anatomic relationships between changes in diffusion-weighted MR image signal intensity, induced by unilateral occlusion of the middle cerebral artery in cats, and tissue perfusion deficits observed in the same animals on T2-weighted MR images after administration of a nonionic intravascular T2 shortening agent. Diffusion-weighted images obtained with strong diffusion-sensitizing gradient strengths (5.6 gauss/cm, corresponding to gradient attenuation factor, b, values of 1413 sec/mm2) displayed increased signal intensity in the ischemic middle cerebral artery territory less than 1 hr after occlusion, whereas T2-weighted images without contrast usually failed to detect injury for 2-3 hr after stroke. After contrast administration (0.5-1.0 mmol/kg by Dy-DTPA-BMA, IV), however, T2-weighted images revealed perfusion deficits (relative hyperintensity) within 1 hr after middle cerebral artery occlusion that corresponded closely to the anatomic regions of ischemic injury shown on diffusion-weighted MR images. Close correlations were also found between early increases in diffusion-weighted MR image signal intensity and disrupted phosphorus-31 and proton metabolite levels evaluated with surface coil MR spectroscopy, as well as with postmortem histopathology. These data indicate that diffusion-weighted MR images more accurately reflect early-onset pathophysiologic changes induced by acute cerebral ischemia than do T2-weighted spin-echo images.