A validation study of the use of near-infrared spectroscopy imaging in primary and secondary motor areas of the human brain.

The electroencephalographically measured Bereitschafts (readiness)-potential in the supplementary motor area (SMA) serves as a signature of the preparation of motor activity. Using a multichannel, noninvasive near-infrared spectroscopy (NIRS) imager, we studied the vascular correlate of the readiness potential. Sixteen healthy subjects performed a self-paced or externally triggered motor task in a single or repetitive pattern, while NIRS simultaneously recorded the task-related responses of deoxygenated hemoglobin (HbR) in the primary motor area (M1) and the SMA. Right-hand movements in the repetitive sequence trial elicited a significantly greater HbR response in both the SMA and the left M1 compared to left-hand movements. During the single sequence condition, the HbR response in the SMA, but not in the M1, was significantly greater for self-paced than for externally cued movements. Nonetheless, an unequivocal temporal delay was not found between the SMA and M1. Near-infrared spectroscopy is a promising, noninvasive bedside tool for the neuromonitoring of epileptic seizures or cortical spreading depolarizations (CSDs) in patients with epilepsy, stroke, or brain trauma because these pathological events are associated with typical spatial and temporal changes in HbR. Propagation is a characteristic feature of these events which importantly supports their identification and characterization in invasive recordings. Unfortunately, the present noninvasive study failed to show a temporal delay during self-paced movements between the SMA and M1 as a vascular correlate of the readiness potential. Although this result does not exclude, in principle, the possibility that scalp-NIRS can detect a temporal delay between different regions during epileptic seizures or CSDs, it strongly suggests that further technological development of NIRS should focus on both improved spatial and temporal resolution. This article is part of a Special Issue entitled Status Epilepticus.

Evaluation of changes in the haemoglobin of skin and muscle tissue of the calf, as induced by topical application of a nonivamide/nicoboxil cream.

Topical agents like nonivamide and nicoboxil induce hyperaemisation and increase cutaneous blood flow and temperature. This study aimed to determine the effects of a nonivamide-nicoboxil cream on haemodynamics in the skin and calf muscle, via optical spectroscopy, discriminating between the changes for skin and muscle. Optical spectroscopy was applied in the visible (VIS) and near-infrared (NIR) wavelength range. The study determined the effect of the cream on changes in oxygenated (ΔoxyHb) and deoxygenated (ΔdeoxyHb) haemoglobin in skin and muscle, as well as on tissue oxygen saturation (SO2) in the skin of 14 healthy subjects. The left and right calves of the subjects were either treated with nonivamide-nicoboxil cream or were sham-administered. NIR spectroscopy allows noninvasive in-vivo examination of the oxygenation of human skeletal muscle. Topical administration of the nonivamide-nicoboxil cream significantly increased the concentration of oxygenated haemoglobin and tissue oxygen saturation in the skin, as well as the concentration of oxygenated haemoglobin in the muscle of the treated legs after 15 min, but with stronger and faster effects in the skin. The topical application of the nonivamide-nicoboxil cream increased blood flow in (smaller vessels of) the skin and muscle tissues.

In the unloaded lower leg, vibration extrudes venous blood out of the calf muscles probably by direct acceleration and without arterial vasodilation.

PURPOSE: During vibration of the whole unloaded lower leg, effects on capillary blood content and blood oxygenation were measured in the calf muscle. The hypotheses predicted extrusion of venous blood by a tonic reflex contraction and that reactive hyperaemia could be observed after vibration. METHODS: Twelve male subjects sat in front of a vibration platform with their right foot affixed to the platform. In four intervals of 3-min duration vibration was applied with a peak-to-peak displacement of 5 mm at frequencies 15 or 25 Hz, and two foot positions, respectively. Near infrared spectroscopy was used for measuring haemoglobin oxygen saturation (SmO2) and the concentration of total haemoglobin (tHb) in the medial gastrocnemius muscle. RESULTS: Within 30 s of vibration SmO2 increased from 55 ± 1 to 66 ± 1 % (mean ± SE). Within 1.5 min afterwards SmO2 decreased to a steady state (62 ± 1 %). During the following 3 min of recovery SmO2 slowly decreased back to base line. THb decreased within the first 30 s of vibration, remained almost constant until the end of vibration, and slowly recovered to baseline afterwards. No significant differences were found for the two vibration frequencies and the two foot positions. CONCLUSIONS: The relaxed and unloaded calf muscles did not respond to vibration with a remarkable reflex contraction. The acceleration by vibration apparently ejected capillary venous blood from the muscle. Subsequent recovery did not match with a reactive hyperaemia indicating that the mere mechanical stress did not cause vasodilation.

Analysis of NIRS-based muscle oxygenation parameters by inclusion of adipose tissue thickness.

The assessment of muscle oxygenation by non-invasive near-infrared spectroscopy generally assumes a homogeneous medium, and this is flawed for large adipose tissue layers underneath the skin. Here we summarize the influence of the adipose tissue thickness on the oxygenation data, show that the adipose layer can be measured by NIRS and indicate a possible correction algorithm. Spectroscopic evidence suggests the usefulness of this algorithm, however, not in all subjects.

Wavelength selection for the improvement of the signal-to-noise ratio for imaging of haemoglobin oxygenation with RGB reflectometry.

We demonstrate the optimisation of wavelengths for the imaging of cortical haemoglobin oxygenation with broadband RGB reflectometry. Wavelengths were chosen in order to minimise the likely crosstalk and optimise the signal-to-noise ratio by simulating effects of different combinations of wavelengths on the condition number of the resulting extinction coefficient matrices. The results obtained were evaluated experimentally for four combinations of commercially available LED combinations and compared with data from the literature.

Tissue oxygenation during exercise measured with NIRS: reproducibility and influence of wavelengths.

Near-infrared spectroscopy (NIRS) is widely used for the measurement of skeletal muscle oxygenation during exercise as it reflects muscle metabolism, and most studies report a large variability between subjects. Here we assess the data quality of tissue oxygen saturation (SO2) and oxygenated (oxyHb) and deoxygenated (deoxyHb) haemoglobin concentrations recorded during an incremental cycling protocol in nine healthy volunteers. The protocol was repeated three times on the same day and a fourth session on a different day to estimate the reproducibility of the method with a broadband, spatially resolved spectroscopy (SRS) system. We found that the inter-subject variation in SO2 (standard deviation ≈ 6 %) was considerably larger than the reproducibility (≈ 1.5 %) both for the same-day and different-day tests. The reproducibility of changes in SO2 was better than 1 %.

Simultaneous imaging of cortical blood flow and haemoglobin concentration with LASCA and RGB reflectometry.

We demonstrate a system for the simultaneous imaging of cortical blood flow and haemoglobin oxygenation by laser speckle contrast analysis (LASCA) and RGB reflectometry. The sensitivity of the system was tested by observing changes of haemoglobin oxygenation and blood flow in rats in response to ischaemic stroke, hypercapnia, hyperoxia, hypoxia, cortical spreading depression and cortical activation following forepaw stimulation.

Cutaneous vasoconstriction affects near-infrared spectroscopy determined cerebral oxygen saturation during administration of norepinephrine.

BACKGROUND: Perioperative optimization of spatially resolved near-infrared spectroscopy determined cerebral frontal lobe oxygenation (scO2) may reduce postoperative morbidity. Norepinephrine is routinely administered to maintain cerebral perfusion pressure and, thereby, cerebral blood flow, but norepinephrine reduces the scO2. We hypothesized that norepinephrine-induced reduction in scO2 is influenced by cutaneous vasoconstriction. METHODS: Fifteen healthy male subjects (25 ± 5 yr, mean ± SD) were studied during: hyperventilation (1.5 kPa end-tidal PcO2 reduction), whole-body heating, administration of norepinephrine (0.15 µg · kg · min; with and without end-tidal carbon dioxide correction), and hypoxia (FiO2: 0.12%). Arterial (saO2), skin, and internal jugular venous oxygen saturations (sjO2) were recorded, and the average cerebral capillary oxygen saturation (scapO2) was calculated. RESULTS: This study indicates that scO2 is influenced by skin oxygen saturation because whole-body heating increased scO2 by 3.6% (2.1-5.1%; 95% CI) and skin oxygen saturation by 3.1% (1.3-4.9%), whereas scapO2 remained unaffected. Conversely, hyperventilation decreased scO2 by 2.1% (0.4-3.7%) and scapO2 by 5.3% (3.8-6.9%), whereas skin oxygen saturation increased 1.8% (0.5-3.1%). In response to hypoxia, scO2 (10.2%; 6.6-13.7%), scapO2 (7.9%; 6.4-9.4%), and skin oxygen saturation (8.9%; 6.3-11.6%) all decreased. With administration of norepinephrine there was a 2.2% (1.0-4.3%) decrease in skin oxygen saturation and scO2 decreased 6.2% (4.2-8.0%), with scapO2 remaining unaffected. CONCLUSION: The results confirm that spatially resolved near-infrared spectroscopy detects cerebral deoxygenation with systemic hypoxic exposure and hyperventilation. However, a commonly used vasopressor norepinephrine disturbs skin oxygen saturation to an extent that influences scO2.

Effects of assuming constant optical scattering on haemoglobin concentration measurements using NIRS during a Valsalva manoeuvre.

Resolving for changes in concentration of tissue chromophores in the human adult brain with near-infrared spectroscopy has generally been based on the assumption that optical scattering and pathlength remain constant. We have used a novel hybrid optical spectrometer that combines multi-distance frequency and broadband systems to investigate the changes in scattering and pathlength during a Valsalva manoeuvre in 8 adult volunteers. Results show a significant increase in the reduced scattering coefficient of 17% at 790nm and 850nm in 4 volunteers during the peak of the Valsalva. However, these scattering changes do not appear to significantly affect the differential pathlength factor and the tissue haemoglobin concentration measurements.

Elevating intracranial pressure reverses the decrease in deoxygenated hemoglobin and abolishes the post-stimulus overshoot upon somatosensory activation in rats.

BOLD fMRI localizes activated brain areas by measuring decreases of deoxygenated hemoglobin (deoxy-Hb) caused by neurovascular coupling. To date, it is unclear whether intracranial pressure (ICP) modifies deoxy-Hb signaling for brain mapping. In addition, ICP elevation can test whether the BOLD post-stimulus undershoot, a transient hypo-oxygenation following functional activation, is due to vascular compliance rather than elevated cerebral metabolic rate of oxygen (CMRO(2)). We addressed these questions by studying the effect of ICP elevation on neurovascular coupling. In anesthetized rats, a cranial window was implanted over the somatosensory cortex. Using laser Doppler flowmetry and optical spectroscopy, changes in cerebral blood flow (CBF), cerebral blood volume (CBV) and deoxy-Hb were measured during electrical forepaw stimulation. Neuronal activity was monitored by somatosensory evoked potentials. ICP was elevated by subarachnoideal and intracisternal infusion of artificial cerebrospinal fluid. ICP elevation did not abrogate neurovascular coupling. However, the concomitant deoxy-Hb decrease was reduced (ICP=14mmHg) and reversed (ICP=28mmHg). Therefore, the validity of BOLD fMRI has to be questioned during increased ICP. Moreover, the amplitude of the deoxy-Hb post-stimulus overshoot was reduced with ICP elevation. CMRO(2) was not elevated during the post-stimulus response. Therefore, these data provide experimental evidence that the BOLD post-stimulus undershoot is a passive vascular phenomenon.

A hybrid multi-distance phase and broadband spatially resolved spectrometer and algorithm for resolving absolute concentrations of chromophores in the near-infrared light spectrum.

For resolving absolute concentration of tissue chromophores in the human adult brain with near-infrared spectroscopy it is necessary to calculate the light scattering and absorption, at multiple wavelengths with some depth resolution. To achieve this we propose an instrumentation configuration that combines multi-distance frequency and broadband spectrometers to quantify chromophores in turbid media by using a hybrid spatially resolved algorithm. Preliminary results in solid phantoms as well as liquid dynamic homogeneous and inhomogeneous phantoms and in-vivo muscle measurements showed encouraging results.

Endothelin-1-induced spreading depression in rats is associated with a microarea of selective neuronal necrosis.

Two different theories of migraine aura exist: In the vascular theory of Wolff, intracerebral vasoconstriction causes migraine aura via energy deficiency, whereas in the neuronal theory of Leão and Morison, spreading depression (SD) initiates the aura. Recently, it has been shown that the cerebrovascular constrictor endothelin-1 (ET-1) elicits SD when applied to the cortical surface, a finding that could provide a bridge between the vascular and the neuronal theories of migraine aura. Several arguments support the notion that ET-1-induced SD results from local vasoconstriction, but definite proof is missing. If ET-1 induces SD via vasoconstriction/ischemia, then neuronal damage is likely to occur, contrasting with the fact that SD in the otherwise normal cortex is not associated with any lesion. To test this hypothesis, we have performed a comprehensive histologic study of the effects of ET-1 when applied topically to the cerebral cortex of halothane-anesthetized rats. Our assessment included histologic stainings and immunohistochemistry for glial fibrillary acidic protein, heat shock protein 70, and transferase dUTP nick-end labeling assay. During ET-1 application, we recorded (i) subarachnoid direct current (DC) electroencephalogram, (ii) local cerebral blood flow by laser-Doppler flowmetry, and (iii) changes of oxyhemoglobin and deoxyhemoglobin by spectroscopy. At an ET-1 concentration of 1 muM, at which only 6 of 12 animals generated SD, a microarea with selective neuronal death was found only in those animals demonstrating SD. In another five selected animals, which had not shown SD in response to ET-1, SD was triggered at a second cranial window by KCl and propagated from there to the window exposed to ET-1. This treatment also resulted in a microarea of neuronal damage. In contrast, SD invading from outside did not induce neuronal damage in the absence of ET-1 (n = 4) or in the presence of ET-1 if ET-1 was coapplied with BQ-123, an ET(A) receptor antagonist (n = 4). In conclusion, SD in presence of ET-1 induced a microarea of selective neuronal necrosis no matter where the SD originated. This effect of ET-1 appears to be mediated by the ET(A) receptor.

Functional imaging with laser speckle contrast analysis: vascular compartment analysis and correlation with laser Doppler flowmetry and somatosensory evoked potentials.

Laser Speckle Contrast Analysis (LASCA), a novel, high-resolution blood flow imaging method, was performed on rat somatosensory cortex during functional activation. In the same animals, cerebral blood flow (CBF) was measured with Laser Doppler Flowmetry. To obtain a quantitative estimate of the underlying neuronal activity, somatosensory evoked potentials were recorded simultaneously with an epidural EEG. Our results show that: 1. CBF changes measured by LASCA or LDF are nonlinearly dependent on the magnitude of electrical neural activity revealed by somatosensory evoked potentials. 2. The magnitude of relative CBF changes measured by LASCA and LDF shows a strong correlation. 3. LASCA imaging localizes the highest relative changes of CBF in microcirculatory areas, with a smaller contribution by larger vessels. This study demonstrates that LASCA is a reliable method that provides 2D-imaging of CBF changes that are comparable to LDF measurements. It further suggests that functional neuroimaging methods based on CBF enhance areas of microcirculation and thus might prove more accurate in localizing neural activity than oxygenation related methods like BOLD-fMRI.

Microcirculation of skeletal muscle adapts differently to a resistive exercise intervention with and without superimposed whole-body vibrations.

Whole-body vibration (WBV) training is commonly practiced and may enhance peripheral blood flow. Here, we investigated muscle morphology and acute microcirculatory responses before and after a 6-week resistive exercise training intervention without (RE) or with (RVE) simultaneous whole-body vibrations (20 Hz, 6 mm peak-to-peak amplitude) in 26 healthy men in a randomized, controlled parallel-design study. Total haemoglobin (tHb) and tissue oxygenation index (TOI) were measured in gastrocnemius muscle (GM) with near-infrared spectroscopy (NIRS). Whole-body oxygen consumption (VO2 ) was measured via spirometry, and skeletal muscle morphology was determined in soleus (SOL) muscle biopsies. Our data reveal that exercise-induced muscle deoxygenation both before and after 6 weeks training was similar in RE and RVE (P = 0.76), although VO2 was 20% higher in the RVE group (P < 0.001). The RVE group showed a 14%-point increase in reactive hyperaemia (P = 0.007) and a 27% increase in blood volume (P < 0.01) in GM after 6 weeks of training. The number of capillaries around fibres was increased by 15% after 6 weeks training in both groups (P < 0.001) with no specific effect of superimposed WBV (P = 0.61). Neither of the training regimens induced fibre hypertrophy in SOL. The present findings suggest an increased blood volume and vasodilator response in GM as an adaptation to long-term RVE, which was not observed after RE alone. We conclude that RVE training enhances vasodilation of small arterioles and possibly capillaries. This effect might be advantageous for muscle thermoregulation and the delivery of oxygen and nutrients to exercising muscle and removal of carbon dioxide and metabolites.

Extra-cerebral oxygenation influence on near-infrared-spectroscopy-determined frontal lobe oxygenation in healthy volunteers: a comparison between INVOS-4100 and NIRO-200NX.

INTRODUCTION: Frontal lobe oxygenation (Sc O2 ) is assessed by spatially resolved near-infrared spectroscopy (SR-NIRS) although it seems influenced by extra-cerebral oxygenation. We aimed to quantify the impact of extra-cerebral oxygenation on two SR-NIRS derived Sc O2 . METHODS: Multiple regression analysis estimated the influence of extra-cerebral oxygenation as exemplified by skin oxygenation (Sskin O2 ) on Sc O2 in 21 healthy subjects exposed to whole-body exercise in hypoxia (Fi O2  = 12%; n = 10) and normoxia (n = 12), whole-body heating, hyperventilation (n = 21), administration of norepinephrine with and without petCO2 -correction (n = 15), phenylephrine and head-up tilt (n = 7). Sc O2 was assessed simultaneously by NIRO-200NX (Sniro O2 ) and INVOS-4100 (Sinvos O2 ). Arterial (Sa O2 ) and jugular bulb oxygen saturations (Sj O2 ) were obtained. RESULTS: The regression analysis indicated that Sinvos O2 reflects 46% arterial, 14% jugular, 35% skin and 4% oxygenation of tissues not interrogated. Sinvos O2 follows a calculated estimate of cerebral capillary oxygenation (r = 0·67; P<0·0001). In contrast, the NIRO-200NX-determined Sc O2 did not correlate with the estimate of cerebral oxygenation (r = 0·026; P = 0·71). CONCLUSION: For all interventions, 35% of the INVOS-4100 signal reflected extra-cerebral oxygenation while, on the other hand, NIRO-200NX did not follow changes in a calculated estimate of cerebral capillary oxygenation. Thus, the NIRO-200NX and INVOS-4100 do not provide for unbiased evaluation of the cerebral signal.

Neuronal activity-induced changes of local cerebral microvascular blood oxygenation in the rat: effect of systemic hyperoxia or hypoxia.

To investigate the effect of resting blood oxygen concentration on the hemodynamic response to functional brain activation, we compared activation-induced changes in hemoglobin oxygenation during normoxia with systemic hyperoxia or mild hypoxia. Hemoglobin oxygenation changes were measured by microfiber optical spectroscopy (500-590 nm) in response to physiological whisker barrel cortex activation by whole whisker pad deflection (4 s, 4 Hz) in alpha-chloralose/urethane anesthetised male Wistar rats. During systemic hyperoxia (n=6), the stimulation-induced hyperoxygenation response was decreased and prolonged, whereas during mild systemic hypoxia (n=7) the peak response was significantly increased followed by a faster return to baseline. During normoxia, a poststimulation under- (oxy-hemoglobin) and overshoot (deoxy-hemoglobin) was observed, which disappeared during systemic hyperoxia and was pronounced during systemic hypoxia. Although averaging out below statistical significance when combining all animals, during mild systemic hypoxia a very small early increase of deoxy-Hb at the beginning of the stimulation period was conspicuous more often than during normoxic or even hyperoxic conditions. This small early increase of deoxy-Hb never preceded the onset of the oxy-Hb response, and was not accompanied by a concomitant decrease in oxy-Hb. Hyperoxia or hypoxia did not affect the induced neuronal responses. Our findings support the concept that the hemodynamic response is regulated according to the metabolic demand of oxygen within the activated tissue.

Noninvasive monitoring of cerebral blood flow by a dye bolus method: separation of brain from skin and skull signals.

Tracking a bolus of contrast agent traveling through the cerebral vasculature provides a measure of the blood flow velocity in the respective cerebral tissue. This principle has been the basis for the first approaches in functional magnetic resonance (MR) imaging and is of great value for investigating patients with vascular disease, especially stroke. While bolus measurements are a standard procedure in MR imaging, optical bolus tracking is as yet not established. Here we study optical absorption changes induced by a bolus of the dye indocyanine-green with near infrared spectroscopy in healthy volunteers. The aim is to assess the latency and shape of the change in absorption. Since application in the adult human critically depends on differentiation between extra- and intracerebral vascular compartments we focus on an approach for such a separation. To do this frequency-domain and multidistance measurements are analyzed by a Monte Carlo based model for photon transport in tissue. Based on measurements of both the photon's mean time of flight (phase) and the intensity, our results allow differentiation between an upper (skin and skull) and a lower layer (brain). The bolus in the deeper tissue layers has a peak of about 10 s width, while the change in absorption in the upper layers shows a much longer recovery time. This is in qualitative agreement with MR imaging results using a gadolinium bolus. This result is promising with respect to the potential of bedside monitoring of mean transit time (MTT) changes in patients with stroke or related vascular disease.

Is leg compression beneficial for alpine skiers?

BACKGROUND: This study examined the effects of different levels of compression (0, 20 and 40 mmHg) produced by leg garments on selected psycho-physiological measures of performance while exposed to passive vibration (60 Hz, amplitude 4-6 mm) and performing 3-min of alpine skiing tuck position. METHODS: Prior to, during and following the experiment the electromygraphic (EMG) activity of different muscles, cardio-respiratory data, changes in total hemoglobin, tissue oxygenation and oscillatory movement of m. vastus lateralis, blood lactate and perceptual data of 12 highly trained alpine skiers were recorded. Maximal isometric knee extension and flexion strength, balance, and jumping performance were assessed before and after the experiment. RESULTS: The knee angle (-10°) and oscillatory movement (-20-25.5%) were lower with compression (P < 0.05 in all cases). The EMG activities of the tibialis anterior (20.2-28.9%), gastrocnemius medialis (4.9-15.1%), rectus femoris (9.6-23.5%), and vastus medialis (13.1-13.7%) muscles were all elevated by compression (P < 0.05 in all cases). Total hemoglobin was maintained during the 3-min period of simulated skiing with 20 or 40 mmHg compression, but the tissue saturation index was lower (P < 0.05) than with no compression. No differences in respiratory parameters, heart rate or blood lactate concentration were observed with or maximal isometric knee extension and flexion strength, balance, and jumping performance following simulated skiing for 3 min in the downhill tuck position were the same as in the absence of compression. CONCLUSIONS: These findings demonstrate that with leg compression, alpine skiers could maintain a deeper tuck position with less perceived exertion and greater deoxygenation of the vastus lateralis muscle, with no differences in whole-body oxygen consumption or blood lactate concentration. These changes occurred without compromising maximal leg strength, jumping performance or balance. Accordingly, our results indicate that the use of lower leg compression in the range of 20-40 mmHg may improve alpine skiing performance by allowing a deeper tuck position and lowering perceived exertion.

Pharmacological uncoupling of activation induced increases in CBF and CMRO2.

Neurovascular coupling provides the basis for many functional neuroimaging techniques. Nitric oxide (NO), adenosine, cyclooxygenase, CYP450 epoxygenase, and potassium are involved in dilating arterioles during neuronal activation. We combined inhibition of NO synthase, cyclooxygenase, adenosine receptors, CYP450 epoxygenase, and inward rectifier potassium (Kir) channels to test whether these pathways could explain the blood flow response to neuronal activation. Cerebral blood flow (CBF) and cerebral metabolic rate of oxygen (CMRO(2)) of the somatosensory cortex were measured during forepaw stimulation in 24 rats using a laser Doppler/spectroscopy probe through a cranial window. Combined inhibition reduced CBF responses by two-thirds, somatosensory evoked potentials and activation-induced CMRO(2) increases remained unchanged, and deoxy-hemoglobin (deoxy-Hb) response was abrogated. This shows that in the rat somatosensory cortex, one-third of the physiological blood flow increase is sufficient to prevent microcirculatory increase of deoxy-Hb concentration during neuronal activity. The large physiological CBF response is not necessary to support small changes in CMRO(2). We speculate that the CBF response safeguards substrate delivery during functional activation with a considerable 'safety factor'. Reduction of the CBF response in pathological states may abolish the BOLD-fMRI signal, without affecting underlying neuronal activity.