Linear aspects of changes in deoxygenated hemoglobin concentration and cytochrome oxidase oxidation during brain activation.

We used near infrared spectroscopy (NIRS) to investigate the vascular and metabolic response to brain activation in human primary and adjacent secondary visual cortex. NIRS is able to measure concentration changes in deoxygenated hemoglobin ([deoxy-Hb]) (which mainly contribute to the blood oxygenation level-dependent (BOLD) signal in functional magnetic resonance imaging (fMRI)) as well as concentration changes of oxygenated hemoglobin ([oxy-Hb]) and corpuscular blood volume ([total-Hb] = [oxy-Hb] + [deoxy-Hb]) and changes in the redox status of the cytochrome c oxidase ([Cyt-Ox]), a putative parameter for cellular oxygenation. A sound understanding of the transfer functions between stimulus parameters, neuronal activity, and vascular/metabolic parameters is important for interpretation of data acquired with indirect neuroimaging techniques like fMRI, especially in event-related design studies. In the present study we tested whether the vascular/metabolic response to stimulation can be described as a linear and time invariant system. Since linearity is a property attributed to systems that satisfy the scaling and superposition properties, as a first simple test, superposition of the responses obtained from short duration visual stimuli was used to predict the responses of longer duration stimuli. Our results showed that the predictions of [deoxy-Hb] and [Cyt-Ox] responses to stimuli of 6- to 24-s duration were satisfactory whereas predictions of [oxy-Hb] and [total-Hb] were insufficient. In a second step, a calculated convolution function of an assumed impulse response function and the stimulus function was fitted with the measured [deoxy-Hb] and [Cyt-Ox] curves to obtain amplitude, time delay, and time constant parameters. We show that predictions of cellular and vascular oxygenation responses to visual stimulation are good for 6- to 24-s stimuli duration under the assumption of a linear transfer characteristic. This model is not valid for corpuscular volume changes which affect mainly the [oxy-Hb] response. Noninvasive NIRS is shown to be a suitable method to get more direct information about neuronal-activity-associated changes in cerebral parameters which are partly reflected in BOLD signal but are not fully understood yet.

Saccadic suppression induces focal hypooxygenation in the occipital cortex.

This study investigated how a decrease in neuronal activity affects cerebral blood oxygenation employing a paradigm of acoustically triggered saccades in complete darkness. Known from behavioral evidence as saccadic suppression, electrophysiologically it has been shown in monkeys that during saccades an attenuation of activity occurs in visual cortex neurons (Duffy and Burchfiel, 1975). In study A, using blood oxygen level-dependent (BOLD) contrast functional magnetic resonance imaging (fMRI), the authors observed signal intensity decreases bilaterally at the occipital pole during the performance of saccades at 2 Hz. In study B.1, the authors directly measured changes in deoxyhemoglobin [deoxy-Hb] and oxyhemoglobin [oxy-Hb] concentration in the occipital cortex with near-infrared spectroscopy (NIRS). Whereas a rise in [deoxy-Hb] during the performance of saccades occurred, there was a drop in [oxy-Hb]. In a second NIRS study (B.2), subjects performed saccades at different rates (1.6, 2.0, and 2.3 Hz). Here the authors found the increase in deoxy-Hb and the decrease of oxy-Hb to be dependent on the frequency of the saccades. In summary, the authors observed a focal hypooxygenation in the human visual cortex dependent on the saccade-frequency in an acoustically triggered saccades paradigm. This could be interpreted as evidence that corresponding to the focal hyperoxygenation observed in functional brain activation, caused by an excessive increase in cerebral blood flow (CBF) over the increase in CMRO2 during decreased neuronal activity CBF, is more reduced than oxygen delivery.

Near-infrared spectroscopy: does it function in functional activation studies of the adult brain?

Changes in optical properties of biological tissue can be examined by near-infrared spectroscopy (NIRS). The relative transparency of tissues including the skull to near-infrared light is the prerequisite to apply the method to brain research. We describe the methodology with respect to its applicability in non-invasive functional research of the adult cortex. A summary of studies establishing the 'typical' response in NIRS vascular parameters, i.e. changes in the concentration of oxygenated and deoxygenated haemoglobin, over an activated area is followed by the validation of changes in the cytochrome-oxidase redox state in response to a visual stimulus. Proceeding from these findings a rough mapping of this metabolic response over the motion-sensitive extrastriate visual area is demonstrated. NIRS measures concentration changes in deoxygenated haemoglobin [deoxy-Hb] which are assumed to be the basis of fMRI BOLD contrast (blood oxygenation level-dependent). The method is therefore an excellent tool to validate assumptions on the physiological basis underlying the fMRI signal, due to its high specificity as to the parameters measured. Questions concerning the concept of 'activation'/'deactivation' and that of the linearity of the vascular response are discussed. To challenge the method we finally present results from a complex single-trial motor paradigm study testing the hypothesis, that premotor potentials (contingent negative variation) can be examined by functional techniques relying on the vascular response. Some of the work described here has been published elsewhere.

Isolation and characterization of circulating 13-kDa C-terminal fragments of human insulin-like growth factor binding protein-5.

The insulin-like growth factor binding proteins (IGFBPs) are responsible for regulation of the effects and the bioavailability of the insulin-like growth factors (IGFs). We screened for circulating fragments of human IGFBP-5 in human hemofiltrate. Identification of IGFBP-5 peptides in the fractions of our peptide bank generated from hemofiltrate was performed by their immunoreactivity and their capacity to bind IGF-I. Different fragments of IGFBP-5 with molecular sizes from 12 to 25 kDa were identified. C-terminal peptides of IGFBP-5 with molecular masses of 13.3 and 13.5 kDa were purified by consecutive chromatographic steps and sequenced. Sequence analysis of the peptides revealed the (double) sequences (K)FVGGAENXAHPRII and MVPRAVYLPNXDRKG. In addition, a smaller fragment with Mr 2722 of the central IGFBP-5 region was purified and showed the sequence HTRISELKAEAVKKDRRKKLTQS (residues 121-143) indicating plasma proteolysis of IGFBP-5 C-terminal to amino acids Lys-120, Ser-143, Lys-144, and Arg-188. According to mass spectrometric and sequence analysis, Thr-152 was shown to be O-glycosylated. Fractions containing C-terminal IGFBP-5 fragments revealed significant IGF-I binding properties. Our results indicate that plasma proteolysis of IGFBP-5 preferentially occurs C-terminally to basic residues and generates different C-terminal fragments, possibly acting in an IGF-dependent manner and bearing intrinsic biological functions.