Assignment (MPhil)--University of Stellenbosch, 2003. / ENGLISH ABSTRACT: The mechanisms of blood flow (BF) control to skeletal muscle during dynamic exercise are still not clearly understood. The paraplegic subject (P) has reduced sympathetic innervation to the lower limbs. The current study was designed to focus on the contribution of neural control, specifically the sympathetic nervous system (SNS), as part of the central vascular mechanism to skeletal muscle BF during dynamic exercise. Aims: We studied BF parameters in P vs. able-bodied subjects (AB) to determine whether the paraplegic can serve as a model for assessing the contribution of the SNS to changes in active vs. inactive muscle BF during exercise. Further questions addressed include: the influence of level of fitness on resting and exercise BF, how lesion level affects BF control in the paraplegic, the 'muscle pump' theory and its hypothesized role in exercise hyperemia and whether blood pooling occurs in the legs of paraplegics. Method: Noninvasive duplex Doppler studies of the large conduit arteries (brachial and common femoral) were performed on 10 elite paraplegic athletes (EP), 10 sedentary paraplegics (SP) en 10 sedentary able-bodied subjects (AB). The paraplegic groups were further subdivided by lesion level with T6 being the critical level. Tests were carried out at rest and after 2 bouts of arm ergometer exercise: a maximal incremental test and 3 minutes at 75% of maximal. Diameter, mean velocity, pulsatile index and blood flow were measured/calculated. Results: Resting heart rate was significantly higher in the paraplegic groups (EP = 80 bpm ± 10, SP = 83 bpm ± 12) vs. the AB group (69 bpm ± 7), p < 0.05. Resting diameter in the common femoral artery (CFA) was similar in EP (5.93 mm ± 1.54) and SP (6.52 mm ± 0.95), but significantly lower than in AB (7.87 mm ± 1.38), p < 0.05. Similar resting pulsatile index (PI) in the CFA were contrary to that previously reported, casting doubt on venous blood pooling theories. Post-exercise values need to be interpreted with caution in view of the large resting differences in CFA diameter. Percentage change values are therefore more appropriate. These differences were not statistically significant, but may suggest interesting trends. Large variability existed for most resting and post-exercise values. Conclusion: The paraplegic subject is an ideal model for the study of the influence of the SNS on blood supply to exercising skeletal muscle. The difference in CFA diameter at rest in the paraplegic vs. the AB group confirms previous results and is probably due to structural/non-physiological changes. Our observation that the BA and CFA diameters in EP and SP subjects do not differ significantly at rest, suggests that training does not have a spillover vasomotor effect on lower limb conduit arteries in paraplegia. Similar BF and PI values post-exercise in the SP and AB groups challenge the muscle pump theory. The SNS has an important role in the control of skeletal muscle blood flow - both at rest (vascular tone) and during exercise (redistribution). Suggestions for future research are made. / AFRIKAANSE OPSOMMING: Die meganismes betrokke by die beheer van bloedvloei (BV) gedurende dinamiese oefening is nog onduidelik. Die parapleeg (P) het verminderde simpatiese innervasie na die onderste ledemate. Die huidige studie fokus op die bydrae van die simpatiese senuwee sisteem (SSS), as deel van die sentrale vaskulêre meganisme, tot skeletale spier BV tydens dinamiese oefening. Doelstellings: Ons het BV parameters in P vs. nie-gestremde proefpersone (kontrole) bestudeer om vas te stelof die parapleeg as model gebruik kan word om die bydrae van die SSS tot veranderings in die BV in aktiewe- en onaktiewe spiere gedurende oefening, te ondersoek. Verdere aspekte wat ondersoek is, sluit in: die invloed van tiksheidvlak ten opsigte van rustende en oefenings BV, of die verlammingsvlak by die parapleeg BV kontrole beïnvloed, die 'spierpomp-teorie' en sy hipotetiese rol in oefeninghiperremie, asook die vraag of bloedsaamstorting in die bene van parapleë plaasvind. Metode: Nie-indringende duplex Doppler studies van die groot geleidingsarteries (bragiaal [BA] en gemene femoral [CFA]) is by 10 elite paraplegiese atlete (EP), 10 sedentêre parapleë (SP) en 10 sedentêre nie-gestremde proefpersone (AB) uitgevoer. Die paraplegiese proefpersone is verder onderverdeel deur die vlak van T6 as kritiese verlammingsvlak te gebruik. Toetse is tydens rus en na 2 arm-ergometer oefeningsessies uitgevoer: een maksimale inkrementeie toets en een van 75% van maksimum intensiteit. Deursnit, gemiddelde vloeispoed, pulsatiewe indeks en bloedvloei is gemeet en/of bereken. Resultate: Rustende hartspoed was beduidend hoër in die paraplegiese groepe (EP = 80 slaelminuut ± 10 en SP = 83 slm ± 12) vs. die AB groep (69 slm ± 7), p < 0.05. Rustende deursnit in die gemene femorale arterie (CFA) was dieselfde in EP (5.93 mm ± 1.54) en SP (6.52 mm ± 0.95), maar beduidend laer as in AB (7.87 mm ± 1.38), p < 0.05. Die feit dat rustende pulsatiewe indeks (PI) in die CFA dieselfde in albei groepe was, laat twyfelontstaan oor die veneuse bloedopdammings teorieë soos weergegee in die literatuur. Na-oefeningswaardes moet omsigtig evalueer word met inagneming van die groot rustende verskille in CFA deursnit. Persentasieverskilwaardes is dus meer toepaslik. Hierdie veskille was nie statisties beduidend nie, maar suggereer interessante tendense. Groot variasie het voorgekom vir beide rustende en na-oefenings waardes. Gevolgtrekking: Die parapleeg is 'n ideale model vir studies om die invloed van die SSS op bloedvloei aan aktiewe skeletale spier te bestudeer. Die verskil in rustende CFA deursnit in die parapleeg vs. die AB groep bevestig vorige resultate en is waarskynlik te wyte aan strukturele, nie-funksionele veranderinge. Ons bevindinge dat die BA en CFA deursneë nie beduidend verskil in die SP en EP groep gedurende rus nie, dui daarop dat gereëlde oefening nie 'n oorloop vasomotor effek op die onderste ledemate in die parapleeg het nie. Die feit dat daar geen verskil aangetoon kon word tussen BV en PI waardes na-oefening in die SP en AB groepe, betwis die spierpomp teorie. Die studie toon dat die SSS 'n belangrike rol in die beheer van skeletale spier bloedvloei speel - beide met rus (vaskulêre tonus) en gedurende oefening (herdistribusie). Voorstelle vir toekomstige navorsing word gemaak.

The hormone glucagon-like peptide-2 (GLP-2) is important in the regulation of intestinal growth and blood flow in nonruminant animals. However, no research reports the existence of GLP-2 in ruminants. Therefore, this dissertation examined the existence of GLP-2 and its receptor, their response to physiological stimuli, and its ability to induce gastrointestinal growth and intestinal blood flow in ruminants. Experiments 1 and 2 established the gastrointestinal distribution of mRNA for proglucagon (the GLP-2 precursor) and the GLP-2 receptor. Furthermore, these experiments determined the effects of changing dietary energy intake on plasma GLP-2 concentrations and proglucagon and GLP-2 receptor mRNA expression. Experiment 3 examined the effect of exogenous bovine GLP-2 on splanchnic blood flow, splanchnic nutrient flux, and gastrointestinal growth. This research shows that ruminants possess a functional GLP-2 signaling system that responds to nutrient ingestion. Based on observed receptor distribution and growth changes with GLP-2 treatment, GLP-2 targets the small intestine and does not affect forestomach or large intestinal growth. Increases in ileal proglucagon mRNA expression and plasma GLP-2 with increasing energy intake demonstrate that GLP-2 responds to physiologic changes in nutrient intake and can be relevant to feeding practices. Furthermore, observed increases in small intestinal growth and blood flow with GLP-2 suggest that it could substantially affect the capacity of the gastrointestinal tract for nutrient absorption. Modification of GLP-2 through diet could allow for improvements in nutrient utilization and animal productivity. This research also has important implications for use of GLP-2 for human disease therapy as the observed downregulation in the blood flow response to 10-d GLP-2 administration has never been reported in any species prior to this dissertation. This research systematically characterized and evaluated the potential role of GLP-2 in the control of gastrointestinal growth and splanchnic blood flow in ruminants. While it extends the knowledge of hormonal control of the gastrointestinal tract in ruminants, it also adds crucial information to the larger body of work investigating the actions of GLP- 2. This dissertation research has contributed to the groundwork necessary to enable the use of GLP-2 in improving the health and productivity of a diverse group of mammalian species.

Ergot alkaloids present in endophyte-infected (E+) tall fescue are thought to be the causative agent of fescue toxicosis, a syndrome affecting cattle in the eastern United States. Many of the observed signs of fescue toxicosis are thought to be attributed to peripheral vasoconstriction; however, there are data indicating that ergot alkaloids can alter blood flow to the gut. An experiment was conducted using right ruminal artery and vein collected from heifers shortly after slaughter. Vessels were mounted in a multi-myograph to determine the vasoconstrictive potentials of ergot alkaloids present in E+ tall fescue. Results indicated ergot alkaloids have the potential to induce vasoconstriction of foregut vasculature. A second experiment was conducted to determine if the additional ergot alkaloids present in E+ tall fescue increase the vasoconstrictive response above that of ergovaline. Results indicated that ergovaline is the main alkaloid responsible for vasoconstriction in bovine vessels. A third study was performed to determine the effect of ergot alkaloids on ruminal epithelial blood flow in the washed rumen of steers exposed to E+ or endophyte-free (E-) tall fescue seed. Steers were dosed with seed followed by a washed rumen experiment with differing levels of ergovaline incubated in the rumen. Results indicated that E+ tall fescue seed treatment reduced ruminal epithelial blood flow. Additionally, incubating ergovaline in the rumen during the washed rumen further decreased epithelial blood flow and volatile fatty acid (VFA) absorption. A final study was conducted to determine the acute effects of ergot alkaloids on isolated rumen epithelial absorptive and barrier functions and the potential for ruminal ergovaline absorption. Results indicate that acute exposure to ergot alkaloids does not alter the absorptive or barrier function of rumen epithelium and ergovaline is absorbed from the rumen. Data from this series of experiments have shown that ergot alkaloids from E+ tall fescue can induce vasoconstriction of blood vessels in the foregut of cattle, reduce blood flow to the rumen epithelium, and decrease VFA absorption. The decrease in nutrient absorption could contribute to the observed symptoms of fescue toxicosis, including depressed growth rates and general unthriftiness.

Atherosclerosis and related cardiovascular diseases (CVDs) are amongst the largest causes of morbidity and mortality in the developed world, causing considerable monetary pressure on public health systems worldwide. Atherosclerosis is characterised by the build up of vascular plaque in medium and large arteries and is a direct precursor to acute vascular syndromes such a myocardial infarction, stroke or peripheral arterial diseases. The causative factors leading to CVD still remain relatively poorly understood, but are becoming increasingly identifiable as a dysfunction of the endothelial cells that line the arterial wall. It is well known that the endothelium responds to the prevailing fluid mechanic (i.e. haemodynamic) environment, which plays a crucial role in the localised occurrence of atherosclerosis near vessel bends and bifurcations. In these areas, disturbed haemodynamics lead to flow separation and very low wall shear stress (WSS), which directly affects the functionality of the endothelium and impedes the transport of important blood borne agonists and antagonists. Detailed full field measurements assessing complex haemodynamics are sparse and consequently this thesis aims to address some of the important questions related to arterial haemodynamics and CVD by performing in-vitro flow measurements in physiologically relevant conditions. In particular, this research develops and uses state-of-the-art Particle Image Velocimetry (PIV) techniques to measure three-dimensional velocity and WSS fields in scaled models of the human carotid artery. For this purpose, the necessary theoretical and experimental concepts are developed and in-depth analyses of the underlying factors affecting the local haemodynamics and their relation to CVD are carried out. In the first part, a methodology for the construct of transparent hydraulic flow phantoms from medical imaging data is developed. The arterial geometries are reproduced in optically clear silicone and the flowing blood is modelled with a refractive index matched blood analogue. Subsequently, planar and Stereo-PIV techniques are developed and verified. A novel interfacial PIV (iPIV) technique is introduced to directly measure WSS by inferring the velocity gradient from the recorded particle images. The new technique offers a maximal achievable resolution of 1 pixel and therefore removes the resolution limit near the wall usually associated with PIV. Furthermore, the iPIV performance is assessed on a number of numerical and experimental test cases and iPIV offers a significantly improved measurement accuracy compared to more traditional techniques. Subsequently, the developed methodologies are applied in three studies to characterise the velocity and WSS fields in the human carotid artery under a number of physiological and experimental conditions. The first study focuses on idealised vessel geometries with and without disease and establishes a general understanding of the haemodynamic environment. Secondly, a physiological accurate vessel geometry under pulsatile flow conditions is investigated to provide a more realistic representation of the true in-vivo flow conditions. The prevailing flow structure in both cases is characterised by flow separation, strong secondary flows and large spatial and temporal variations in WSS. Large spatial and temporal differences exist between the different geometries and flow conditions; spatial variations appear to be more significant than transient events. Thirdly, the three-dimensional flow structure in the physiological carotid artery model is investigated by means of stereoscopic and tomographic PIV, permitting for the first time the measurement of the full 3D-3C velocity field and shear stress tensor in such geometries. The flow field within the model is complex and three-dimensional and inherently determined by the vessel geometry and the build up of an adverse pressure gradient. The main features include strong heliocoidal flow motions and large spatial variations in WSS. Lastly, the physiological implications of the current results are discussed in detail and reference to previous work is given. In summary, the present research develops a novel and versatile PIV methodology for haemodynamic in vitro studies and the functionality and accuracy is demonstrated through a number of physiological relevant flow measurements.

For simulations of highly complex geometries, frequently encountered in many fields of science and engineering, the process of generating a high-quality, body-fitted grid is very complicated and time-intensive. Thus, one of the principal goals of contemporary CFD is the development of numerical algorithms, which are able to deliver computationally efficient, and highly accurate solutions for a wide range of applications involving multi-physics problems, e.g. Fluid Structure Interaction (FSI). Immersed interface/boundary methods provide considerable advantages over conventional approaches, especially for flow problems containing moving boundaries.In the present work, a novel, robust, highly-accurate, Immersed Interface Method (IIM) is developed, which is based on a local Taylor-series expansion at irregular grid points enforcing numerical stability through a local stability condition. Various immersed methods have been developed in the past; however, these methods only considered the order of the local truncation error. The numerical stability of these schemes was demonstrated (in a global sense) by considering a number of different test-problems. None of these schemes used a concrete local stability condition to derive the irregular stencil coefficients. This work will demonstrate that the local stability constraint is valid as long as the DFL-number does not reach a limiting value. The IIM integrated into a newly developed Incompressible Navier-Stokes (INS) solver is used herein to simulate fully coupled FSI problems. The extension of the novel IIM to a higher-order method, the compressible Navier-Stokes equations and the Maxwell's equations demonstrate the great potential of the novel IIM.In the second part of this dissertation, the newly developed INS solver is employed to study the flow of a stalled airfoil and steady/unsteady stenotic flows. In this context, a new biglobal stability analysis approach based on solving an Initial Value Problem (IVP), instead of the traditionally used EigenValue Problem (EVP), is presented. It is demonstrated that this approach based on an IVP is computationally less expensive compared to EVP approaches while still capturing the relevant physics.

<p>Optical coherence tomography (OCT) is a non-invasive imaging modality that uses low coherence interferometry to generate three-dimensional datasets of a sample's structure. OCT has found tremendous clinical applications in imaging the retina and has demonstrated great utility in the diagnosis of various retinal diseases. However, such diagnoses rely upon the ability to observe abnormalities in the structure of the retina caused by pathology. By the time an ocular disease has progressed to the point of affecting the morphology of the retina, irreversible vision loss in the eye may already occur. Changes in the functionality of the tissue often precede changes to the structure. Thus, if imaging methods are developed to provide additional functional information about the behavior and response of the retinal tissue and vasculature, earlier treatment for disease may be prescribed, thus preserving vision for the patient. </p><p>Within the last decade, significant technological advances in OCT systems have enabled high-speed and high sensitivity image acquisition using either spectral domain OCT (SDOCT) or swept-source OCT (SSOCT) configurations. Such systems use Fourier processing to extract structural information of a sample from interferometric principles. But such systems also have access to the optical phase information, which allows for functional analysis of sample dynamics. This dissertation details the development and application of methods using both intensity and phase information as a tool for studying interesting biological phenomena. The goal of this work is an extension of techniques to image the vasculature in the retina and enhance the clinical utility of OCT.</p><p>I first outline basic theory necessary for understanding the principles of OCT. I then describe OCT phase imaging in cellular applications as a demonstration of the ability of OCT to provide functional information on biological dynamics. Phase imaging methods suffer from an artifact known as phase wrapping, and I have developed a software technique to overcome this problem in OCT, thus extending its usefulness in providing quantitative information. I characterize the limitations in measuring moving scatterers with Doppler OCT in both SDOCT and SSOCT system. I also show the ability to image the vasculature in the retina using variance imaging with a high-speed retinal imaging system and software based methods to correct for patient motion and create a widefield mosaic in an automated manner. Finally, future directions for this work are discussed.</p> / Dissertation

The presence of an alpha-adrenoceptor--mediated coronary vasoconstrictor reflex during acute systemic hypoxia was examined in thirteen chloralose-anesthetized dogs. Local vasodilator effects were avoided by perfusing the left common coronary artery (LCC) with normoxic blood, while the dogs were ventilated with 5% 02-95% N2 . Left ventricular afterload was held constant and positive cardiac inotropic responses and beta two-adrenoceptor-mediated coronary vasodilation were blocked by propranolol. Parasympatheticmediated bradycardia and coronary vasodilation were blocked with atropine. Systemic hypoxia decreased LCC flow to normoxic myocardium by 19.4+2.6 %. Although myocardial oxygen extraction increased 9.7+2.9 %, myocardial oxygen consumption decreased 16.5+2.6 %. Intracoronary prazosin prevented the reflex vasoconstriction during repeated hypoxia.

Introduction and aim: Increases in the bioavailability of nitric oxide have been shown to reduce the oxygen (O2) cost of exercise, improve exercise performance, alter gastric blood flow and mediate glucose uptake in healthy individuals. Aim: Does dietary nitrate reduce the O2 cost of exercise, improve walking performance in individuals with type 2 diabetes mellitus (T2DM) and chronic obstructive pulmonary disease (COPD) or alter hepatic diffusion and positively affect glucose homeostasis in healthy adults? Methods: Experimental chapters utilised a double-blind, placebo-controlled, randomised, experimental design. Breath by breath pulmonary gas analysis was utilised to assess the O2 cost of exercise in 48 individuals with T2DM and 13 with COPD. Walking performance was assessed via the six minute walk test (6MWT) in cohorts 1 and 2. Magnetic resonance imaging was used to assess portal vein flux, velocity and the apparent diffusion coefficient, in order to assess hepatic microvascular diffusion (apparent diffusion coefficient (ADC)). Blood pressure (BP) was measured in all trials. Results: Relative to placebo, beetroot juice resulted in a significant increase in plasma nitrate and nitrite. There were no differences between placebo vs. beetroot juice for the O2 cost of walking (T2DM: placebo; 946 ± 221 vs. beetroot juice; 939 ± 223 ml.min-1; P=0.59) or cycling (COPD: placebo; 933 ± 323 vs. beetroot juice; 939 ± 302 ml: min-1; P=0.88), distance covered in the 6MWT (T2DM: placebo; 550 ± 83 vs. beetroot juice; 554 ± 90m; P=0.17 or COPD: placebo; 456 ± 86 vs. beetroot juice; 449 ± 79 m; P=0.37) or BP (T2DM: systolic: placebo; 134 ± 10 vs. beetroot juice; 132 ± 12 mmHg, P=0.17; diastolic: placebo; 77 ± 7: vs. beetroot juice; 76 ± 11 mmHg, P=0.27. COPD: systolic: placebo; 123 ± 14 vs. beetroot juice; 123 ± 14 mmHg; P=0.91; diastolic: placebo; 77 ± 9 vs. beetroot juice; 79 ± 9 mmHg; P=0.27). No differences were seen between placebo and beetroot juice for ADC (young adults: F(3, 45) = 0.25, P = 0.74; older adults; F(3, 42) = 1.3, P = 0.28), portal vein flux (young adults: F(3, 45) = 0.339, P = 0.79; older adults; F(3, 42) = 1.65, P = 0.19) however, there was an interaction effect in the young adults: (F(3, 45) = 2.9, P = 0.04) but not in the older adults; F(3, 42) = 1.8, P = 0.16) between visits for portal vein velocity. Nitrate supplementation did not reduce plasma glucose concentrations (young adults: F(3, 45) = 0.96, P = 0.42; older adults; F(3, 42) = 0.04, P = 0.99). Nitrate supplementation did not reduce systolic blood pressure (young adults: F(3, 45) = 0.20, P = 0.89; older adults; F(3, 42) = 1.7, P = 0.18) or diastolic blood pressure (young adults: F(3, 45) = 0.25, P = 0.86; older adults; F(3, 42) = 0.45, P = 0.72). Conclusion Dietary nitrate supplementation does not alter the O2 cost of exercise, improve walking performance or reduce BP in individuals with T2DM or COPD. Nitrate supplementation does not alter hepatic diffusion, glucose homeostasis or BP.

A number of investigators have examined psychological stress-induced endothelial dysfunction, however, the underlying mechanisms for these responses have not been clearly elucidated. The purpose of this study was to compare the effects of mental challenge on forearm blood flow, total antioxidant capacity (a measure of oxidative stress), the release of norepinephrine (NE; stress induced neurotransmitter), and pro-inflammatory cytokine responses [both lipopolysaccharide (LPS)-stimulated TNF-α and IL-6 cytokine and mRNA] in lean and obese individuals. Twelve subjects who had a BMI above 30 kg/m2 and were above 30% body fat were categorized as obese and twelve subjects with a BMI below 25 kg/m2 and were below 25% body fat were categorized as lean subjects. Blood samples were drawn and forearm blood flow was assessed prior to and following subjects’ participation in a mental challenge protocol consisting of a computer-based Stroop Color-Word task and mental arithmetic task, for a total of 20 minutes. The mental challenge elicited an elevation in HR and NE in both the lean and obese groups. Furthermore, both lean and obese groups demonstrated an increase in FBF following the mental challenge, whereas no changes in total antioxidant capacity were observed. In addition, the lean group exhibited an increase in LPS-stimulated TNF-α cytokine production from baseline to following the mental challenge, whereas the obese group demonstrated a decrease in LPS-stimulated TNF-α cytokines. This corresponded with a decrease in LPS-stimulated TNF-α mRNA expression in the obese group, although the obese subjects maintained higher levels of both measurements (LPS-stimulated TNF-α cytokine and mRNA expression) compared with the lean group. Furthermore, in the LPS-stimulated IL-6 cytokine response, the obese group demonstrated a greater increase than the lean group following the mental challenge, even though both groups showed an increase in LPS-stimulated IL-6 mRNA expression. These findings suggest that the magnitude and direction of LPS-stimulated TNF-α cytokine response and mRNA expression and LPS-stimulated IL-6 cytokine response to acute stress may be dependent upon the effects of the additional percentage of body fat seen in obese individuals.

In resting muscle, blood flow is regulated to meet the demand for O2 by the tissue. A modified ischemia (I)/reperfusion(R) investigation was systematically run and PISFO2, PaO2, Q and VO2 were observed. Twenty-nine spinotrapezius muscles from male Sprague-Dawley rats (284±20 grams) were surgically exteriorized for intravital microscopy to test a model relating blood flow, O2 supply and O2 demand. The model can aid in the understanding of the regulation of tissue PO2. The interstitial PO2 (PISFO2) and perivascular PO2 (PaO2) measurements were made using phosphorescence quenching microscopy (PQM). O2 consumption (VO2) values were obtained with a quasi-continuous, flash-synchronized, pressurized airbag to initiate ischemia and sample the rate of O¬2 change (dPO2/dt). Centerline red blood cell velocity was measured with an Optical Doppler Velocimeter and converted to flow using vessel diameter. 5-, 15-, 30-, 60-, 300- and 600-second ischemic durations were used to observe changes in PISFO2, Q, and VO2. A critical point was observed following 30 seconds of (I) where dPISFO2/dt during recovery was the fastest (4.25±0.72 mmHg/s) and was 1.00±0.16 mmHg/s following 600 seconds. Flow recovery, dQ/dt, peaked to 3.88±0.64 (µl•min-1)/s after 60 seconds of (I) but significantly dropped to 2.83±0.55 (µl•min-1)/s following 300 seconds of (I) but increased to 2.92±0.45 (µl•min-1)/s following 600 seconds. This gives evidence to a no-reflow phenomenon occurring in the extended periods of ischemia. A peak in VO¬2 to 309.2±45.0 nl O2/cm3•s with a time course of 160 seconds occurred following 600 seconds of ischemia. As the ischemic duration decreased, the time course and peak VO2 also decreased. VO2 following 300 seconds of (I) was significantly higher than 5-60 seconds of (I) (p <0.05) but was not significantly different from 600 seconds of (I). The information collected during the Q and VO2 studies can be incorporated into a factor, M, that relates VO2, Q and ∆PO2. M calculated for the recovery of 5- through 60-second (I) groups reasonably relates the three variables due to consistency and little variability. However, recovery in 600- and especially 300-second (I) groups showed higher variability in M which requires more consideration.