Thirty minutes of handgrip exercise enhances brachial artery flow-mediated dilation in response to two different shear stress profiles.

The walls of blood vessels are lined with a single-cell layer of endothelial cells. As blood flows through the arteries, a frictional force known as shear stress is sensed by mechanosensitive structures on the endothelium. Short and long term changes in shear stress can have a significant influence on the regulation of endothelial function. Acutely, shear stress triggers a pathway that culminates in the release of vasodilatory molecules from the endothelium and subsequent vasodilation of the artery. This endothelial response is known as flow mediated dilation (FMD). FMD is used as an index of endothelial function and is commonly assessed using reactive hyperemia (RH)-FMD, a method which elicits a large, short lived increase in shear stress following the release of a brief (5 min) forearm occlusion. A recent study found that a short term exposure (30 min) to a sustained elevation in shear stress potentiates subsequent RH-FMD. FMD can also result from a more prolonged, sustained increase in shear stress elicited by handgrip exercise (HGEX-FMD). There is evidence to suggest that interventions and conditions impact FMD resulting from sustained and transient shear stress stimuli differently, indicating that HGEX-FMD and RH-FMD provide different information about endothelial function. It is unknown whether HGEX-FMD is improved by short term exposure to shear stress. Understanding how exercise induced FMD is regulated is important because it contributes to blood flow responses during exercise. The study purpose was therefore to assess the impact of a handgrip exercise (intervention) induced sustained elevation in shear stress on subsequent brachial artery (BA) HGEX-FMD. Twenty healthy male participants (22±3yrs) preformed a 30-minute HGEX intervention on two experimental days. BA-FMD was assessed using either an RH or HGEX shear stress stimulus at 3 time points: pre-intervention, 10 min post and 60 min post. FMD and shear stress magnitude were determined via ultrasound. Both HGEX and RH-FMD increased significantly from pre-intervention to 10 min-post (p<0.01). These findings indicate that FMD stimulated by exercise induced increases in shear stress is potentiated by short term shear stress exposure. These findings advance our understanding regarding the regulation of endothelial function by shear stress.

Does estrogen fluctuation throughout the menstrual cycle impact flow-mediated dilation during exercise in healthy premenopausal women?

The endothelium is the inner most layer of cells that lines all arteries. A primary function of endothelial cells is to regulate responses to increased blood flow and the resulting frictional forces or shear stress by producing factors such as nitric oxide that mediate arterial dilation (flow mediated dilation (FMD)). Menstrual cycle variations in estrogen (E2) have been shown to influence brachial artery (BA) FMD in response to transient increases in shear stress brought about by the release of a brief forearm occlusion (reactive hyperemia (RH)). FMD can also be assessed in response to a sustained shear stress stimulus such as that created with handgrip exercise (HGEX), and studies have shown that RH- and HGEX stimulated FMD provide unique information regarding endothelial function. However, the impact of menstrual phase on HGEX-FMD is unknown. Therefore, the purpose of this study was to determine the impact of cyclical changes in E2 levels on HGEX-FMD over two discrete phases of the menstrual cycle. FMD was assessed via ultrasound. 12 subjects (21 ± 2yrs) completed two experimental visits: (1) low estrogen phase (early follicular) and (2) High estrogen phase (late follicular). In each visit both RH- and HGEX-FMD (6 min handgrip exercise) were assessed. Results are mean ± SD. E2 increased from the low to the high estrogen phase of the menstrual cycle (low: 34 ± 8, high: 161 ± 113pg/mL, p = 0.004). There was no change in mean FMD between phases (RH-FMD: 7.7 ± 4.3% vs. 6.4 ± 3.1%, p = 0.139; HGEX-FMD: 4.8 ± 2.8% vs. 4.8 ± 2.3%, p = 0.979). The observation that both RH- and HGEX-FMD did not differ between phases indicates that menstrual cycle fluctuations in estrogen may not universally impact endothelial function in young, healthy premenopausal women. Further research is needed to improve our understanding of the mechanisms that underlie variability in the impact of menstrual phase on both transient and sustained FMD responses.

A Quantitative and Mechanistic Assessment of Activated Thrombin-Activatable Fibrinolysis Inhibitor and its Role in Pathological Bleeding and Thrombosis

The coagulation and fibrinolytic systems are linked by the thrombin-thrombomodulin complex which regulates each system through activation of protein C and TAFI, respectively. We have used novel assays and techniques to study the enzymology and biochemistry of TAFI and TAFIa, to measure TAFI activation in hemophilia A and protein C deficiency and to determine if enhancing TAFI activation can improve hemostasis in hemophilic plasma and whole blood. We show that TAFIa not TAFI attenuates fibrinolysis in vitro and this is supported by a relatively high catalytic efficiency (16.41μM-1s-1) of plasminogen binding site removal from fibrin degradation products (FDPs) by TAFIa. Since the catalytic efficiency of TAFIa in removing these sites is ~60-fold higher than that for inflammatory mediators such as bradykinin it is likely that FDPs are a physiological substrate of TAFIa. The high catalytic efficiency is primarily a result of a low Km which can be explained by a novel mechanism where TAFIa forms a binary complex with plasminogen and is recruited to the surface of FDPs. The low Km also suggests that TAFIa would effectively cleave lysines from FDPs during the early stages of fibrinolysis (i.e. at low concentrations of FDPs). Since individuals with hemophilia suffer from premature fibrinolysis as a result of insufficient TAFI activation we quantified TAFI activation in whole blood from hemophilic subjects. Both the rate of activation and the area under the TAFI activation time course (termed TAFIa potential) was determined to be reduced in hemophilia A and the TAFIa potential was significantly and inversely correlated with the clinical bleeding iii phenotype. Using a novel therapeutic strategy, we used soluble thrombomodulin to increase TAFI activation which improved the clot lysis time in factor VIII deficient human plasma and hemophilic dog plasma as well as hemophilic dog blood. Finally, we briefly show in a biochemical case study that TAFI activation is enhanced in protein C deficiency and when afflicted individuals are placed on Warfarin anticoagulant therapy, TAFI activation is reduced. Since TAFIa stabilizes blood clots, this suggests that reducing TAFI activation or inhibiting TAFIa may help restore blood flow in vessels with pathological thrombosis.

Higher brain neurons succumb to acute stroke-like insult while lower brain neurons strongly resist

Pyramidal neurons (PyNs) in ‘higher’ brain are highly susceptible to acute stroke injury yet ‘lower’ brain regions better survive global ischemia, presumably because of better residual blood flow. Here we show that projection neurons in ‘lower’ brain regions of hypothalamus and brainstem intrinsically resist acute stroke-like injury independent of blood flow in the brain slice. In contrast `higher` projection neurons in neocortex, hippocampus, striatum and thalamus are highly susceptible. In live brain slices from rat deprived of oxygen and glucose (OGD), we imaged anoxic depolarization (AD) as it propagates through these regions. AD, the initial electrophysiological event of stroke, is a depolarizing front that drains residual energy in compromised gray matter. The extent of AD reliably determines ensuing damage in higher brain, but using whole-cell recordings we found that all CNS neurons do not generate a robust AD. Higher neurons generate strong AD and show no functional recovery in contrast to neurons in hypothalamus and brainstem that generate a weak and gradual AD. Most dramatically, lower neurons recover their membrane potential, input resistance and spike amplitude when oxygen and glucose is restored, while higher neurons do not. Following OGD, new recordings could be acquired in all lower (but not higher) brain regions, with some neurons even withstanding multiple OGD exposure. Two-photon laser scanning microscopy confirmed neuroprotection in lower, but not higher gray matter. Specifically pyramidal neurons swell and lose their dendritic spines post-OGD, whereas neurons in hypothalamus and brainstem display no such injury. Exposure to the Na+/K+ ATPase inhibitor ouabain (100 μM), induces depolarization similar to OGD in all cell types tested. Moreover, elevated [K+]o evokes spreading depression (SD), a milder version of AD, in higher brain but not hypothalamus or brainstem so weak AD correlates with the inability to generate SD. In summary, overriding the Na+/K+ pump using OGD, ouabain or elevated [K+]o evokes steep and robust depolarization of higher gray matter. We show that this important regional difference can be largely accounted for by the intrinsic properties of the resident neurons and that Na+/K+ ATPase pump efficiency is a major determining factor generating strong or weak spreading depolarizations.