High-Resolution Time-Frequency Analysis Of Neurovascular Responses To Ischemic Challenges

The identification and delineation of oscillatory patterns of blood flow associated with cerebral autoregulation mechanisms is the focus of three specific aims. 1) Identify a superior spectral analysis method for the identification of activity due to cholinergic oscillatory control of the microvasculature (COCmicvasc) and compare the effectiveness of the Hilbert-Huang transform (HHT), the smoothed pseudo Wigner-Ville (SPWV) distribution, and the variable-frequency complex demodulation (VFCDM) method to detect such activity. 2) Determine if changes in forehead microcirculation as monitored by noninvasive laser Doppler flowmetry may provide a reliable indication of the adequacy of cerebral blood flow during progressive simulated hypovolemia. 3) Identify changes in forehead microvascular activity that is due to parasympathetic activity associated with the onset of mental status changes and show a lack of coherence with potential confounding variables. The lower body negative pressure (LBNP) protocol is a noninvasive technique that can induce a hypovolemic state through blood pooling in the lower extremities by a portion of the circulating blood volume. Increasing degrees of LBNP is applied to healthy volunteers being monitored with noninvasive laser Doppler (LD) flowmetry to assess levels of forehead perfusion and vascular resistance. MATLAB is utilized as the primary tool for the development of custom mathematical algorithms to analyze this data in the time-frequency-energy domain. Utilizing the VFCDM technique to analyze forehead laser Doppler data offers greater insight into the identification and understanding of cerebral autoregulatory mechanisms than using other available methods. Techniques to identify COCmicvasc activity in the forehead and distinguish it from respiration, a potential confounder, are discussed. Presented data provides evidence that the observed COCmicvasc activity is due to a locally induced autoregulatory mechanism in the forehead that is unrelated to passive transmission of the mechanical effects of respiration. The close relationship of the forehead vasculature to that of the brain (i.e. part of the forehead is fed by a branch of the internal carotid artery with the remainder from the external carotid artery) suggests that the observed autoregulatory processes in the forehead microvasculature may be the same mechanisms responsible for autoregulation of the brain and other vital organs. This conclusion is also supported by the correlation between changes in mental status (i.e. lightheadedness) and forehead COCmicvasc activity

Detection of Spatial and Temporal Interactions in Renal Autoregulation Dynamics

Renal autoregulation stabilizes renal blood flow to protect the glomerular capillaries and maintain glomerular filtration rates through two mechanisms: tubuloglomerular feedback (TGF) and the myogenic response (MR). It is considered that the feedback mechanisms operate independently in each nephron (the functional unit of the kidney) within a kidney, but renal autoregulation dynamics can be coupled between vascular connected nephrons. It has also been shown that the mechanisms are time-varying and interact with each other. Understanding of the significance of such complex behavior has been limited by absence of techniques capable of monitoring renal flow signals among more than 2 or 3 nephrons simultaneously. The purpose of this thesis was to develop approaches to allow the identification and characterization of spatial and temporal properties of renal autoregulation dynamics. We present evidence that laser speckle perfusion imaging (LSPI) effectively captures renal autoregulation dynamics in perfusion signals across the renal cortex of anaesthetized rats and that spatial heterogeneity of the dynamics is present and can be investigated using LSPI. Next, we present a novel approach to segment LSPI of the renal surface into phase synchronized clusters representing areas with coupled renal autoregulation dynamics. Results are shown for the MR and demonstrate that when a signal is present phase synchronized regions can be identified. We then describe an approach to identify quadratic phase coupling between the TGF and MR mechanisms in time and space. Using this approach we can identify locations across the renal surface where both mechanisms are operating cooperatively. Finally, we show how synchronization between nephrons can be investigated in relation to renal autoregulation effectiveness by comparing phase synchronization estimates from LSPI with renal autoregulation system properties estimated from renal blood flow and blood pressure measurements. Overall, we have developed approaches to 1) capture renal autoregulation dynamics across the renal surface, 2) identify regions with phase synchronized renal autoregulation dynamics, 3) quantify the presence of the TGF-MR interaction across the renal surface, and 4) determine how the above vary over time. The described tools allow for investigations of the significance and mechanisms behind the complex spatial interactions and time-varying properties of renal autoregulation dynamics

Linear and Nonlinear Modeling of Cerebral Flow Autoregulation Using Principal Dynamic Modes

Cerebral Flow Autoregulation (CFA) is the dynamic process by which cerebral blood flow is maintained within physiologically acceptable bounds during fluctuations of cerebral perfusion pressure. The distinction is made with “static” flow autoregulation under steady-state conditions of perfusion pressure, described by the celebrated “autoregulatory curve” with a homeostatic plateau. This paper studies the dynamic CFA during changes in perfusion pressure, which attains critical clinical importance in patients with stroke, traumatic brain injury and neurodegenerative disease with a cerebrovascular component. Mathematical and computational models have been used to advance our quantitative understanding of dynamic CFA and to elucidate the underlying physiological mechanisms by analyzing the relation between beat-to-beat data of mean arterial blood pressure (viewed as input) and mean cerebral blood flow velocity(viewed as output) of a putative CFA system. Although previous studies have shown that the dynamic CFA process is nonlinear, most modeling studies to date have been linear. It has also been shown that blood CO2 tension affects the CFA process. This paper presents a nonlinear modeling methodology that includes the dynamic effects of CO2 tension (or its surrogate, end-tidal CO2) as a second input and quantifies CFA from short data-records of healthy human subjects by use of the modeling concept of Principal Dynamic Modes (PDMs). The PDMs improve the robustness of the obtained nonlinear models and facilitate their physiological interpretation. The results demonstrate the importance of including the CO2 input in the dynamic CFA study and the utility of nonlinear models under hypercapnic or hypocapnic conditions

Sodium channel regulatory mechanisms : current fluctuation analysis on frog skin epithelium

This project examined the role of the cytoskeleton in regulatory mechanisms of the amiloride-sensitive Na⁺ channels in isolated frog skin epithelium. The epithelium from ventral frog skin is a model tissue which has proved significant in our understanding of the basic principles involved in water and Na⁺ homeostasis. In particular, this project examines ways in which local (non-hormonal) and hormonal regulatory mechanisms adjust the Na⁺ permeability of apical membranes of frog skin epithelium. Both mechanisms contain factors that are known to increase the apical membrane Na⁺ permeability mainly by increases in the number of open channels. The origin of these new open channels is unknown but, it is postulated that they could arise either by activation of quiescent channels already present in the apical membrane, or by recruitment of channels from cytoplasmic stores. Regarding the latter hypothesis, we also examined the idea that the cytoskeleton might somehow be involved in the insertion of Na⁺ channels within vesicles, into the apical membrane. This is based on the fact that the cytoskeleton is involved in a similar mechanism whereby, in the toad urinary bladder, anti-diuretic hormone (ADH) causes the insertion of aggregates with water channels. Much current interest focuses on the role of the cytoskeleton in the regulation of epithelial Na⁺ channels. To test this hypothesis, we used noise analysis to examine the effects of disrupting the cytoskeleton, on two different mechanisms which bring about changes in open channel densities. The mechanisms are: (1) lowering mucosal Na⁺ concentration (non-hormonal), and (2) addition of arginine-vasopressin (A VP) (hormonal). Non-hormonal, autoregulatory changes in apical membrane Na⁺ conductance were examined by investigating the effects of reducing the mucosal Na⁺ concentration. Our results showed that lowering the mucosal Na⁺ concentration induced large increases in the open channel density in order to stabilise the transport rate. In addition, we observed an average 55-60% increase in the open channel probability, which implies that in epithelium from Rana fuscigula, changes of channel open probability are also an important mechanism in the autoregulation of channel densities in response to a reduction in mucosal Na⁺. The hormonal control of Na⁺ channels by A VP has been intensively studied by noise analysis and the patch clamp. Our results confirmed previous reports that A VP increases the Na⁺ transport rate by increasing the number of open Na⁺ channels, primarily through large changes in the total number of channels, without a significant change in open probability. Regarding the role of the cytoskeleton in regulation of Na⁺ channels and/or its possible role in control of inserting putative vesicles with Na⁺ channels, we studied the effects of disrupting the cytoskeleton on the two regulatory mechanisms. Disrupting microtubules with colchicine had no, or very little effect on either of the regulatory mechanisms. On the other hand, the integrity of the microfilaments was very important for the autoregulatory changes in the number of open channels. After cytochalasin B treatment, lowering the mucosal Na⁺ concentration did not result in the usual compensatory changes in channel densities. There was no prior evidence that cytochalasin B had any actual effect on the F-actin network in the frog skin epithelium. Accordingly, modified cytochemical techniques were designed to demonstrate and localise F-actin in the epithelial granular cells. The direct immunofluorescent method proved useful, but did not allow sufficient resolution to examine the changes to different populations of actin in the cells. We then modified an immunogold method to suit our conditions, and the results demonstrated the localisation of different pools of F-actin and showed the effects of the cytochalasin B and vasopressin

Forehead laser doppler and transcranial doppler during simulated hypovolemia

The present study employed lower body negative pressure (LBNP), a rapidly titratable, safe and reversible means of inducing simulated hypovolemia, for a comparison of transcranial Doppler (TCD) ultrasound of the middle cerebral artery and laser Doppler (LD) flowmetry of the forehead microvasculature. With IRB approval, 9 healthy volunteers (26.3±2.7 years) were monitored continuously with EKG, noninvasive finger arterial blood pressure (BP), and TCD positioned at the transtemporal window. After a baseline (Base) period, subjects underwent rapid onset of LBNP to -70 mmHg over the course of 1 minute, followed by progressive declines of ~10 mmHg until lightheadedness or had a BP decline \u3e20% of baseline BP. Changes in the peak (systolic) and trough (diastolic) values with each heart beat were analyzed at Base, at approx. 30 seconds prior to the onset of lightheadedness (Presympt) and at onset of symptoms (Sympt). In the 6 subjects who subsequently became lightheaded, forehead LD flow decreased by 10.9±11.7% at Presympt (p=NS for interphase difference). It then decreased by an additional 20.4±18.7% with the onset of lightheadedness (p=0.035 for Presympt vs. Sympt). Peak TCD readings decreased by 29.3±9.7% from Base to the time of the Presympt measurement (p=0.001); they then increased by 4.1±12.9% with the onset of Sympt (p=NS). In the 2 subjects who remained asymptomatic, LD did not change significantly in the Presympt and Sympt phases where Sympt was the time when the study was terminated because the BP cutoff was reached. In these asymptomatic subjects, the TCD flow velocity declined progressively. The present findings suggest that monitoring of the microvasculature in the distribution of the carotid arteries provides a better indication of changes in perfusion associated with lightheadedness than measurement of velocity at the middle cerebral artery. The discordance between LD and TCD is consistent with autoregulatory mechanisms at the level of the forehead microvasculature that have previously been reported in the context of systemic administration of phenylephrine

On the Indeterminates of Glaucoma:the Controversy of Arterial Blood Pressure and Retinal Perfusion

Glaucoma is a chronic eye disease characterized by thinning of the retina, death of ganglion cells, and progressive loss of vision, eventually leading to blindness. The prevalence of glaucoma is estimated at 1-3% of those over 40 years old. With a constantly aging population, this number is expected to increase significantly over the next 10 years. Even with treatment, about 15% of people with glaucoma currently develop residual vision or tunnel vision and eventually become blind or partially sighted. The mechanisms behind ganglion cell death are poorly understood. Elevated eye pressure is the main risk factor for glaucoma, but treatment in the form of medication, laser, or surgery can only slow the decline, not stop it. In addition, high intraocular pressure is neither necessary nor sufficient for the development of glaucoma, indicating the existence of other unknown risk factors. It has been established that the death of ganglion cells results in a decreased oxygen demand and a concomitant decrease in blood flow. However, there is also a hypothesis that reduced or unstable blood supply is not only a consequence, but also a cause of glaucoma. This is known as the ‘chicken-egg’ dilemma in glaucoma. It is supported by the observation that the risk of developing glaucoma is higher in people with very low blood pressure (sometimes even as a result of overtreatment of high blood pressure).This dissertation is an attempt to methodically examine whether blood pressure can be linked to changes in the retina that could suggest susceptibility to glaucoma. For this purpose, we analyze epidemiological data from the Groningen Longitudinal Glaucoma Study, we use advanced imaging techniques to model the microcirculation, and we describe its relationship with the neural structure and oxygen consumption of the retina. We provide evidence leaning towards the existence of a vascular component, likely pertinent to glaucoma

Dynamics of Erythropoietic Survival Pathways In Vivo: A Dissertation

Erythropoiesis maintains stable tissue oxygenation in the basal state, while accelerating red cell production in anemia, blood loss or high altitude. The principal regulator of erythropoiesis is the hormone erythropoietin (Epo). In response to hypoxic stress, Epo can increase a 1000-fold, driving erythropoietic rate by up to 10-fold. It’s been suggested that survival pathways activated by the Epo receptor (EpoR) underlie its regulation of erythropoietic rate. A number of apparently redundant EpoR survival pathways were identified in vitro, raising the possibility of their functional specialization in vivo. Here I assessed the roles of three survival pathways activated by EpoR in erythroblasts in-vivo: the suppression of cell-surface Fas and FasL, the suppression of the pro-apoptotic regulator Bim, and the induction of the anti-apoptotic regulator Bcl-xL. I used the novel CD71/Ter119 flow-cytometric method of identifying erythroblast maturation stages in vivo to measure these apoptotic pathways in fetal liver and adult erythropoietic tissues. I found that these pathways differ markedly in their regulation of erythropoietic rate. Using mouse genetic models, I found that apoptosis mediated by interaction between erythroblasts that co-express cell-surface Fas and FasL plays a key autoregulatory role in stabilizing the size of the erythroblast pool in the basal state. Further, mice mutant for Fas or FasL showed a delayed erythropoietic response to hypoxia or high Epo. This suggests that Fas and FasL accelerate the stress response by providing an apoptotic ‘cell reserve’ that can be rescued by Epo in stress. I also examined the in-vivo behavior of two cell-intrinsic apoptotic regulators, Bcl-xL and Bim, previously unexamined in stress. The induction of Bcl-xL was rapid but transient, whilst the suppression of Bim was slower but persistent. My data suggest that Bcl-xL is a key mediator of EpoR’s anti-apoptotic signal very early in the stress response, before Bim and Fas are suppressed. Bcl-xL adaptation to high Epo occurs through inhibition of Stat5 activation, and resets it for the next acute stress. My findings suggest that in vivo, Epo regulates erythropoietic rate through erythroblast apoptosis, and that various apoptotic regulators play distinct and unique roles in this process. My work provides new molecular insights into erythropoiesis that are relevant to cytokine biology and to clinical approaches of disease treatment

Aerospace Medicine and Biology: A continuing bibliography (supplement 160)

This bibliography lists 166 reports, articles, and other documents introduced into the NASA scientific and technical information system in October 1976