Daisuke Furuya

Diffuse Optical Tomography of Cerebral Blood Flow, Oxygenation, and Metabolism in Rat during Focal Ischemia

Diffuse optical tomography (DOT) is an attractive approach for evaluating stroke physiology. It provides hemodynamic and metabolic imaging with unique potential for continuous noninvasive bedside imaging in humans. To date there have been few quantitative spatial-temporal studies of stroke pathophysiology based on diffuse optical signatures. The authors report DOT images of hemodynamic and metabolic contrasts using a rat middle cerebral artery occlusion (MCAO) stroke model. This study used a novel DOT device that concurrently obtains coregistered images of relative cerebral blood volume (rCBV), tissue-averaged hemoglobin oxygen saturation (Sto2), and relative cerebral blood flow (rCBF). The authors demonstrate how these hemodynamic measures can be synthesized to calculate an index of the oxygen extraction fraction (OEF) and the cerebral metabolic rate of oxygen consumption (CMRo2). Temporary (60-minute) MCAO was performed on five rats. Ischemic changes, averaged over the 60 minutes of occlusion, were as follows: rCBF = 0.42 ± 0.04, rCBV = 1.02 ± 0.04, ΔSto2 = −11 ± 2%, rOEF = 1.39 ± 0.06 and rCMRo2 = 0.59 ± 0.07. Although rOEF increased in response to decreased blood flow, rCMRo2 decreased. The sensitivity of this method of DOT analysis is discussed in terms of assumptions about baseline physiology, and the diffuse optical results are compared with positron emission tomography, magnetic resonance imaging, and histology observations in the literature.

A Peroxisome Proliferator-Activated Receptor-γ Agonist Reduces Infarct Size in Transient but not in Permanent Ischemia

Background and Purpose— Activators of peroxisome proliferator-activated receptor-&ggr; (PPAR&ggr;), a member of the PPAR family, increase levels of CuZn-superoxide dismutase (SOD) in cultured endothelium, suggesting a mechanism by which it may exert its protective effect within the brain. These properties raise the question of whether a PPAR&ggr; agonist may be neuroprotective in models of ischemia without reperfusion, in which oxidative injury is less prevalent. Methods— In 2 groups of rats, 90 minutes of middle cerebral artery (MCA) occlusion was followed by 1 day of reperfusion, with 1 group receiving pioglitazone (a PPAR&ggr; agonist) starting 72 hours before MCA occlusion (MCAO) and continuing through the day of occlusion, whereas the other group received vehicle only. In 2 comparable groups, the MCA was occluded permanently. One day after occlusion, the animals were tested neurologically and infarct volumes were calculated. In a separate group, rats were treated with pioglitazone or vehicle for 4 days. Tissue was obtained from the cortex and the striatum 2 hours into reperfusion after 90 minutes of MCAO, and the tissue was examined for CuZn-SOD by Western blot. Results— Results show a significant reduction in infarct size in the treated rats, with transient MCAO but not permanent MCAO. There was also an improvement in neurological score in the treated animals after transient MCAO. The level of CuZn-SOD was increased in the cortex in treated animals. Conclusions— These data, which show that a PPAR&ggr; agonist reduces infarct size in transient but not permanent MCAO, suggest that the role of PPAR&ggr; is specific to events occurring during reperfusion. Our data point to CuZn-SOD as the mediator of this neuroprotection.

Spatiotemporal Quantification of Cerebral Blood Flow during Functional Activation in Rat Somatosensory Cortex using Laser-Speckle Flowmetry

Laser-speckle flowmetry was used to characterize activation flow coupling after electrical somatosensory stimulation of forepaw and hindpaw in the rat. Quantification of functional activation was made with high transverse spatial (μm) and temporal (msec) resolution. Different activation levels and duration of stimulation were quantitatively investigated, and were in good agreement with previous laser-Doppler measurements. Interestingly, the magnitude but not the overall shape of the response was found to scale with stimulus amplitude and the distance from the activation centroid. The results provide new insights about the spatial characteristics of cerebral blood flow response to functional activation, and the method should lead to improved understanding of the coupling of neuronal activity and hemodynamics under normal and pathologic conditions.

Diffuse optical correlation tomography of cerebral blood flow during cortical spreading depression in rat brain

Diffuse optical correlation methods were adapted for three-dimensional (3D) tomography of cerebral blood flow (CBF) in small animal models. The image reconstruction was optimized using a noise model for diffuse correlation tomography which enabled better data selection and regularization. The tomographic approach was demonstrated with simulated data and during in-vivo cortical spreading depression (CSD) in rat brain. Three-dimensional images of CBF were obtained through intact skull in tissues(~4mm) deep below the cortex.

Frequency-domain multiplexing system for in vivo diffuse light measurements of rapid cerebral hemodynamics

A novel frequency-domain multiplexing system has been developed for in vivo measurements of rapid cerebral hemodynamics. The instrument operates in the frequency-domain with three optical wavelengths, six source positions, and two detectors. Frequency-division multiplexing was used to modulate three wavelengths (690, 786, and 830 nm) at slightly different frequencies around 70 MHz. The three laser output beams were combined and switched into different source positions by use of fast optical switches (switch time <10 ms). Three narrowband, in-phase and in-quadrature demodulators decode the modulated signals. Our full-frame-acquisition rate is 2.5 Hz, with flexibility for acquisition rates greater than 50 Hz with smaller detection areas. We evaluate the performance of the instrument with tissue phantoms, and then employ the system to measure in vivo cerebral blood oxygenation during forepaw stimulation of a rats brain.

Diffuse Optical Measurement of Hemoglobin and Cerebral Blood Flow in Rat Brain During Hypercapnia, Hypoxia and Cardiac Arrest

We have demonstrated the ability to concurrently measure relative changes in cerebral blood flow, hemoglobin concentration, and hemoglobin oxygenation with a single non-contact, non-invasive instrument. Our measurements from rat hypercapnia, hypoxia and cardiac arrest models are in reasonable agreement with the literature, and offer the possibility for further growth and quantification. The optical techniques used in this study are attractive also because they enable experimenters to measure vascular response of deep tissues. The new instrument and concept may also be applicable to human studies especially in infants and neonates permitting noninvasive monitoring of cerebral hemodynamics and oxygen (see [4] and [2] for examples of NIR spectroscopy).

Rats recovering from unilateral barrel–cortex ischemia are capable of completing a whisker-dependent task using only their affected whiskers

Rats use their vibrissae for a variety of exploratory tasks including location of objects and discrimination of texture. This study examines recovery in vibrissal function following a unilateral ischemic injury to the somatosensory cortex. Vibrissal function was examined in adult food-restricted rats performing on a two-texture discrimination device. Animals were trained and tested until the criteria of >80% correct choices was demonstrated on three consecutive days. Ischemic rats were constrained to use the affected whiskers by clipping the ipsilateral vibrissae. One group was tested after ischemia, a second group was trained before ischemia and then tested, and a third group was pre-trained and received whisker stimulation and tested post-ischemia. Nai;ve animals recovering from ischemia took longer to reach criteria than intact or unilateral trimmed control animals. Pre-trained animals with compression ischemia receiving whisker stimulation with sucrose water completed the task to criteria in the fewest number of trials. The results indicate that recovery of vibrissal function occurs following a unilateral ischemic injury. Histological analysis in animals without whisker stimulation indicates that the number of normal appearing cortical barrels following ischemia was inversely correlated to the number of trials needed to complete the behavioral task. This suggests that the natural recovery of the ability to discriminate textures is related to the degree of damage to the barrel cortex. The relationship between cortical barrels and behavioral recovery did not hold for the ischemic animals receiving whisker stimulation. This latter group demonstrated recovery despite marked anatomical lesions suggesting that the intervention influenced reorganization.

Effects of telmisartan on the cerebral circulation of hypertensive patients with chronic-stage stroke

This prospective study examined the effects of telmisartan, an angiotensin II type I receptor blocker with peroxisome proliferator-activated receptor gamma agonistic action, on blood pressure (BP) control and cerebral circulation in hypertensive patients with chronic-stage stroke. Telmisartan (40 mg per day) was administered to 10 patients with systolic BP (SBP) ⩾140 mm Hg and diastolic BP (DBP) ⩾90 mm Hg at least 4 weeks after lacunar or atherothrombotic infarction. Casual BP and resting cerebral blood flow (CBF) were evaluated at baseline and week 12 using technetium-99 m ethyl cysteinate dimer single-photon emission computed tomography. Both SBP and DBP declined significantly from 156.4±17.0 to 127.4±6.6 mm Hg and 84.2±14.5 to 74.2±5.2 mm Hg, respectively (P<0.05). Mean CBF (mCBF) in both the left and right cerebral hemispheres did not change, and the mCBF of both the impaired and unimpaired sides of supratentorial lesion patients (n=6) did not change. Investigation of regional CBF in all patients revealed significant increases in the callosomarginal, precentral, central, parietal, temporal, posterior cerebral, lenticular nucleus, thalamic and hippocampal regions at week 12 (P<0.05). Telmisartan showed good antihypertensive activity in hypertensive patients with chronic-stage stroke without affecting hemispheric blood flow, and it even increased regional CBF in most regions examined.

Spatio-temporal quantification of cerebral blood flow during forepaw stimulation of the rat using laser speckle flowmetry

Laser speckle flowmetry was used to characterize activation flow coupling following forepaw/hindpaw stimulation. Different activation levels and duration were investigated. Temporal response of activation area was independent of stimulus amplitude and distance from the centroid.

Hemodynamic measurements in rat brain and human muscle using diffuse near-infrared absorption and correlation spectroscopies

Measurement of concentration, oxygenation, and flow characteristics of blood cells can reveal information about tissue metabolism and functional heterogeneity. An improved multifunctional hybrid system has been built on the basis of our previous hybrid instrument that combines two near-infrared diffuse optical techniques to simultaneously monitor the changes of blood flow, total hemoglobin concentration (THC) and blood oxygen saturation (StO2). Diffuse correlation spectroscopy (DCS) monitors blood flow (BF) by measuring the optical phase shifts caused by moving blood cells, while diffuse photon density wave spectroscopy (DPDW) measures tissue absorption and scattering. Higher spatial resolution, higher data acquisition rate and higher dynamic range of the improved system allow us to monitor rapid hemodynamic changes in rat brain and human muscles. We have designed two probes with different source-detector pairs and different separations for the two types of experiments. A unique non-contact probe mounted on the back of a camera, which allows continuous measurements without altering the blood flow, was employed to in vivo monitor the metabolic responses in rat brain during KCl induced cortical spreading depression (CSD). A contact probe was used to measure changes of blood flow and oxygenation in human muscle during and after cuff occlusion or exercise, where the non-contact probe is not appropriate for monitoring the moving target. The experimental results indicate that our multifunctional hybrid system is capable of in vivo and non-invasive monitoring of the hemodynamic changes in different tissues (smaller tissues in rat brain, larger tissues in human muscle) under different conditions (static versus moving). The time series images of flow during CSD obtained by our technique revealed spatial and temporal hemodynamic changes in rat brain. Two to three fold longer recovery times of flow and oxygenation after cuff occlusion or exercise from calf flexors in a patient with peripheral vascular disease (PVD) were found.