Charity L. Sauder

Aging affects the cardiovascular responses to cold stress in humans.

Cardiovascular-related mortality peaks during cold winter months, particularly in older adults. Acute physiological responses, such as increases in blood pressure, in response to cold exposure may contribute to these associations. To determine whether the blood pressure-raising effect (pressor response) of non-internal body temperature-reducing cold stress is greater with age, we measured physiological responses to 20 min of superficial skin cooling, via water-perfused suit, in 12 younger [25 +/- 1 (SE) yr old] and 12 older (65 +/- 2 yr old) adults. We found that superficial skin cooling elicited an increase in blood pressure from resting levels (pressor response; P < 0.05) in younger and older adults. However, the magnitude of this pressor response (systolic and mean blood pressure) was more than twofold higher in older adults (P < 0.05 vs. younger adults). The magnitude of the pressor response was similar at peripheral (brachial) and central (estimated in the aorta) measurement sites. Regression analysis revealed that aortic pulse wave velocity, a measure of central arterial stiffness obtained before cooling, was the best predictor of the increased pressor response to superficial skin cooling in older adults, explaining approximately 63% of its variability. These results indicate that there is a greater pressor response to non-internal body temperature-reducing cold stress with age in humans that may be mediated by increased levels of central arterial stiffness.

Effects of posture on shear rates in human brachial and superficial femoral arteries

Shear rate is significantly lower in the superficial femoral compared with the brachial artery in the supine posture. The relative shear rates in these arteries of subjects in the upright posture (seated and/or standing) are unknown. The purpose of this investigation was to test the hypothesis that upright posture (seated and/or standing) would produce greater shear rates in the superficial femoral compared with the brachial artery. To test this hypothesis, Doppler ultrasound was used to measure mean blood velocity (MBV) and diameter in the brachial and superficial femoral arteries of 21 healthy subjects after being in the supine, seated, and standing postures for 10 min. MBV was significantly higher in the brachial compared with the superficial femoral artery during upright postures. Superficial femoral artery diameter was significantly larger than brachial artery diameter. However, posture had no significant effect on either brachial or superficial femoral artery diameter. The calculated shear rate was significantly greater in the brachial (73 +/- 5, 91 +/- 11, and 97 +/- 13 s(-1)) compared with the superficial femoral (53 +/- 4, 39 +/- 77, and 44 +/- 5 s(-1)) artery in the supine, seated, and standing postures, respectively. Contrary to our hypothesis, our current findings indicate that mean shear rate is lower in the superficial femoral compared with the brachial artery in the supine, seated, and standing postures. These findings of lower shear rates in the superficial femoral artery may be one mechanism for the higher propensity for atherosclerosis in the arteries of the leg than of the arm.

Neurovascular responses to mental stress in the supine and upright postures

The purpose of this study was to determine neurovascular responses to mental stress (MS) in the supine and upright postures. MS was elicited in 23 subjects (26 +/- 1 yr) by 5 min of mental arithmetic. In study 1 (n = 9), Doppler ultrasound was used to measure mean blood flow velocity in the renal (RBFV) and superior mesenteric arteries (SMBFV), and venous occlusion plethysmography was used to measure forearm blood flow (FBF). In study 2 (n = 14), leg blood flow (LBF; n = 9) was measured by Doppler ultrasound, and muscle sympathetic nerve activity (MSNA; n = 5) was measured by microneurography. At rest, upright posture increased heart rate and MSNA and decreased LBF, FBF, RBFV, and SMBFV and their respective conductances. MS elicited similar increases in mean arterial blood pressure ( approximately 12 mmHg) and heart rate ( approximately 17 beats/min), regardless of posture. MS in both postures elicited a decrease in RBFV, SMBFV, and their conductances and an increase in LBF, FBF, and their conductances. Changes in blood flow were blunted in the upright posture in all vascular beds examined, but the pattern of the vascular response was the same as the supine posture. MS did not change MSNA in either posture (change: approximately 1 +/- 3 and approximately 3 +/- 3 bursts/min, respectively). In conclusion, the augmented sympathetic activity of the upright posture does not alter heart rate, mean arterial blood pressure, or MSNA responses to MS. MS elicits divergent vascular responses in the visceral and peripheral vasculature. These results indicate that, although the upright posture attenuates vascular responses to MS, the pattern of neurovascular responses does not differ between postures.

Melatonin differentially affects vascular blood flow in humans

Melatonin is synthesized and released into the circulation by the pineal gland in a circadian rhythm. Melatonin has been demonstrated to differentially alter blood flow to assorted vascular beds by the activation of different melatonin receptors in animal models. The purpose of the present study was to determine the effect of melatonin on blood flow to various vascular beds in humans. Renal (Doppler ultrasound), forearm (venous occlusion plethysmography), and cerebral blood flow (transcranial Doppler), arterial blood pressure, and heart rate were measured in 10 healthy subjects (29±1 yr; 5 men and 5 women) in the supine position for 3 min. The protocol began 45 min after the ingestion of either melatonin (3 mg) or placebo (sucrose). Subjects returned at least 2 days later at the same time of day to repeat the trial after ingesting the other substance. Melatonin did not alter heart rate and mean arterial pressure. Renal blood flow velocity (RBFV) and renal vascular conductance (RVC) were lower during the melatonin trial compared with placebo (RBFV, 40.5±2.9 vs. 45.4±1.5 cm/s; and RVC, 0.47±0.02 vs. 0.54±0.01 cm·s(-1)·mmHg(-1), respectively). In contrast, forearm blood flow (FBF) and forearm vascular conductance (FVC) were greater with melatonin compared with placebo (FBF, 2.4±0.2 vs. 1.9±0.1 ml·100 ml(-1)·min(-1); and FVC, 0.029±0.003 vs. 0.023±0.002 arbitrary units, respectively). Melatonin did not alter cerebral blood flow measurements compared with placebo. Additionally, phentolamine (5-mg bolus) after melatonin reversed the decrease in RVC, suggesting that melatonin increases sympathetic outflow to the kidney to mediate renal vasoconstriction. In summary, exogenous melatonin differentially alters vascular blood flow in humans. These data suggest the complex nature of melatonin on the vasculature in humans.

Mental Stress Elicits Sustained and Reproducible Increases in Skin Sympathetic Nerve Activity.

Mental stress (MS) is a known trigger of myocardial infarction and sudden death. By activating the sympathetic nervous system, MS may have deleterious effect on the cardiovascular system but this process is not completely understood. The primary aim of this study was to quantify the effect of MS on skin sympathetic nerve activity (SSNA). The secondary aim was to determine the reproducibility of SSNA to MS within a given day and ~1 week later. Ten subjects (26 ± 1 year) performed two bouts of mental arithmetic lasting 3 min. The bouts were separated by 45 min. One week later the subjects returned to repeat MS. All experiments were conducted in the supine posture during the morning hours. To maintain neutral skin temperature, each subject wore a custom suit (34–35°C). Skin blood flow and sweat rate were measured on the dorsal foot. MS elicited a marked increase in SSNA within the first 10 sec (184 ± 42%; P < 0.01) in all subjects, which was less during the remaining period of MS, but remained elevated (87 ± 20; P < 0.01). The pattern of responses to MS was unchanged during the second bout (10 sec, 247 ± 55%; 3 min average, 133 ± 29%) and during the retest 1 week later (10 sec, 196 ± 55%; 3 min average, 117 ± 36%). MS did not significantly affect cutaneous vascular conductance or sweat rate during any trial. In summary, MS elicits robust and reproducible increases in SSNA in humans, which may be followed over time to observe alterations in the regulation of the autonomic nervous system.

Aging attenuates the vestibulorespiratory reflex in humans

Activation of the vestibular system changes ventilation in humans. The purpose of the present study was to investigate whether aging alters the vestibulorespiratory reflex in humans. Because aging attenuates the vestibulosympathetic reflex, it was hypothesized that aging would attenuate the vestibulorespiratory reflex. Changes in ventilation during engagement of the semicircular canals and/or the otolith organs were measured in fourteen young (26 ± 1 years) and twelve older subjects (66 ± 1 years). In young subjects, natural engagement of the semicircular canals and the otolith organs by head rotation increased breathing frequency during dynamic upright pitch at 0.25 Hz (15 cycles min−1) and 0.5 Hz (30 cycles min−1) (Δ2 ± 1 and Δ4 ± 1 breaths min−1, respectively; P < 0.05) and during dynamic upright roll (Δ2 ± 1 and Δ4 ± 1, respectively; P < 0.05). In older subjects, the only significant changes in breathing frequency occurred during dynamic pitch and roll at 0.5 Hz (Δ2 ± 1 and Δ2 ± 1 for pitch and roll, respectively). Stimulation of the horizontal semicircular canals by yaw rotation increased minute ventilation in young but not older subjects. Selective engagement of the otolith organs during static head‐down rotation did not alter breathing frequency in either the young or older subjects. The results of this study indicate that the vestibulorespiratory reflex is attenuated in older humans, with greater vestibular stimulation needed to activate the reflex.

Melatonin attenuates the skin sympathetic nerve response to mental stress

Melatonin attenuates muscle sympathetic nerve responses to sympathoexcitatory stimuli, but it is unknown whether melatonin similarly attenuates reflex changes in skin sympathetic nerve activity (SSNA). In this double-blind, placebo-controlled, crossover study, we tested the hypothesis that melatonin (3 mg) would attenuate the SSNA response to mental stress (mental arithmetic). Twelve healthy subjects underwent experimental testing on two separate days. Three minutes of mental stress occurred before and 45 min after ingestion of melatonin (3 mg) or placebo. Skin temperature was maintained at 34°C. Reflex increases in SSNA (peroneal nerve), mean arterial pressure, and heart rate (HR) to mental stress before and after melatonin were determined. Melatonin lowered HR (pre, 66 ± 3 beats/min; and post, 62 ± 3 beats/min, P = 0.046) and SSNA (pre, 14,282 ± 3,706 arbitrary units; and post, 9,571 ± 2,609 arbitrary units, P = 0.034) at rest. In response to mental stress, SSNA increases were significantly attenuated following melatonin ingestion (second minute, 114 ± 30 vs. 74 ± 14%; and third minute, 111 ± 29 vs. 54 ± 12%, both P < 0.05). The mean arterial pressure increase to mental stress was blunted in the third minute (20 ± 2 vs. 17 ± 2 mmHg, P = 0.032), and the HR increase was blunted in the first minute (33 ± 3 vs. 29 ± 3 beats/min, P = 0.034) after melatonin. In summary, exogenous melatonin attenuates the SSNA response to mental stress.

Greater sensitivity of the vestibulosympathetic reflex in the upright posture in humans

Otolith organs have been shown to activate the sympathetic nervous system in the prone position by head-down rotation (HDR) in humans. To date, otolithic stimulation by HDR has not been comprehensively studied in the upright posture. The purpose of the present study was to determine whether otolithic stimulation increases muscle sympathetic nerve activity (MSNA) in the upright posture. It was hypothesized that stimulation of the otolith organs would increase MSNA in the upright posture, despite increased baseline sympathetic activation due to unloading of the baroreceptors. MSNA, arterial blood pressure, heart rate, and degree of head rotation were measured during HDR in 18 volunteers (23 +/- 1 yr) in different postures. Study 1 (n = 11) examined HDR in the prone and sitting positions and study 2 (n = 7) examined HDR in the prone and 60 degrees head-up tilt positions. Baseline MSNA was 8 +/- 4, 15 +/- 4, and 33 +/- 2 bursts/min for prone, sitting, and head-up tilt, respectively. HDR significantly increased MSNA in the prone (Delta4 +/- 1 and Delta105 +/- 37% for burst frequency and total activity, respectively), sitting (Delta5 +/- 1 and Delta43 +/- 12%), and head-up tilt (Delta7 +/- 1 and Delta110 +/- 41%; P < 0.05). Sensitivity of the vestibulosympathetic reflex (%DeltaMSNA/DeltaHDR; degree of head rotation) was significantly greater in the sitting and head-up tilt than prone position (prone = 74 +/- 22; sitting = 109 +/- 30; head-up tilt = 276 +/- 103; P < 0.05). These data indicate that stimulation of the otolith organs can mediate increases in MSNA in the upright posture and suggest a greater sensitivity of the vestibulosympathetic reflex in the upright posture in humans.

Interactive effect of aging and local muscle heating on renal vasoconstriction during isometric handgrip

The purpose of the study was to determine the interactive effect of aging and forearm muscle heating on renal vascular conductance and muscle sympathetic nerve activity (MSNA) during ischemic isometric handgrip. A tube-lined, water-perfused sleeve was used to heat the forearm in 12 young (27 +/- 1 yr) and 9 older (63 +/- 1 yr) subjects. Ischemic isometric handgrip was performed before and after heating. Muscle temperature (intramuscular thermistor) was 34.3 +/- 0.2 and 38.7 +/- 0.1 degrees C during normothermia and heating, respectively. At rest, heating had no effect on renal blood velocity (Doppler ultrasound) or renal vascular conductance in either group (young, n = 12; older, n = 8). Heating compared with normothermia caused a significantly greater increase in renal vasoconstriction during exercise and postexercise muscle ischemia (PEMI) in both groups. However, the increase in renal vasoconstriction during heating was greater in the older compared with the young subjects (18 +/- 3 vs. 8 +/- 3%). During handgrip, heating elicited greater increases in MSNA responses in the older group (young, n = 12; older, n = 6), whereas no statistical difference was observed between groups during PEMI. In summary, aging augments renal vascular responses to ischemic isometric handgrip during heating of the exercising muscle. The greater renal vasoconstriction was associated with augmented MSNA in the older subjects.

Aerobic Training Improves In Vivo Cholinergic Responsiveness but Not Sensitivity of Eccrine Sweat Glands

TO THE EDITOR Aerobic training improves thermal tolerance (Armstrong and Pandolf, 1988), which is postulated to result in part from improved evaporative cooling (Taylor, 1986; Shibasaki et al., 2006). Previous observations indicate that aerobic training either lowers the internal temperature at which sweating begins (sweating threshold) or increases the slopes of the internal temperaturesweat rate (Roberts et al., 1977) or exercise intensity–sweat rate relations (Yanagimoto et al., 2002). In contrast, deconditioning or detraining, associated with bedrest, increases the sweating threshold and decreases the slope of internal temperature–sweat rate relation (Lee et al., 2002). Importantly these deconditioning-related responses can be prevented by exercising during bedrest (Shibasaki et al., 2003). These studies, although informative, do not provide mechanistic insight into whether altered sweating responses to aerobic training are mediated by a central sympathetic component or at the level of the eccrine gland. Application of cholinergic agonists directly in the dermal space without repeated injections or electric current can isolate peripheral sweating responses (Crandall et al., 2003; Morgan et al., 2006; Schlereth et al., 2006). Cross-sectional studies (comparing aerobic trained vs. untrained) using electrical current drug delivery (iontophoresis) have observed increased sweating capacity and number of activated sweat glands with aerobic training (Buono and Sjoholm, 1988; Buono et al., 1992). However, these studies provided minimal insight into whether these adaptatory responses were mediated by changes in receptor responsiveness and sensitivity, or were simply a function of subject population selected. Accordingly, we tested the hypothesis that 8 weeks of aerobic training increases in vivo cholinergic sensitivity and responsiveness of eccrine sweat glands, without altering the number of exogenous acetylcholineactivated glands. Eleven sedentary, young (age1⁄4 26±1), healthy, non-obese (BMIo 30 kg m), normotensive (o140/90 mm Hg), non-smokers participated in this institutional approved longitudinal training study that adhered to Declaration of Helsinki guidelines. Each subject provided written informed consent and received a medical history and physical exam before participating in the study. Two intradermal microdialysis membranes were placed B3 cm apart in dorsal forearm skin in a similar location preand post-training, which unlike previous studies allows for continuous monitoring of sweat glands at a prescribed agonist concentration (Morgan et al., 2006). This technique involved placing a small (200-mm outer diameter, 10-mm length) semipermeable membrane (20-kDa cutoff) intradermally at a depth of approximately 0.3–1.0 mm below the epidermis (Kellogg et al., 1999). Eight doses of acetylcholine (10 7 to 1 M acetylcholine) in a lactated Ringer’s vehicle were administered for 5 minutes at 2 ml/minutes via a microdialysis infusion pump. Sweat rate was measured via capacitance hygrometry and the number of activated sweat glands was quantified using a starch–iodine technique during infusion of 1 M acetylcholine. Cholinergic dose–response relations were determined by logistic regression modeling. Endurance training consisted of running or cycling 4 times/week for 8 weeks. Subjects wore a heart rate monitor during all exercise sessions to ensure maintenance of target heart rates. Training began with exercising for 20 minutes at a work rate sufficient to achieve 80% of maximum heart rate. Exercise times increased to 60 minutes as training progressed, and high-intensity interval exercises were added twice/week during the second week to further stimulate training adaptations. Peak oxygen uptake increased from 32.9 to 40.7 ml kg 1 min 1 or 19±2% (Po0.05) and resting heart rate decreased 9±2 b.p.m. (Po0.05) indicative of a training effect. The number of cholinergic-activated glands was unchanged by training (pre-training1⁄4 40±6 and post-training1⁄4 39±7 glands per 0.5 cm). This indicates that a similar number of glands were recruited both preand post-training and that this type and duration of training does not increase the number of exogenous cholinergic-activated glands. Dose–response relations were established with goodness of fit (R) relations of 0.67±0.02 for pre-training and 0.72±0.02 for post-training, indicating that the logistic regression modeling adequately modeled the data (Figure 1). Training did not affect the ED50, but increased the maximal responses of the dose–response relation (Figure 2). These findings provide important insight into the adaptatory mechanism(s) of eccrine sweat glands to longitudinal aerobic training in humans. First, we did not observe a leftward shift in the ED50 of the cholinergic dose–response curve, strongly suggesting that exercise training does not alter eccrine sweat gland in vivo cholinergic sensitivity as previously suggested (Roberts et al., 1977; Buono et al., 1992). However, these reports suggesting an increase in cholinergic sensitivity used an iontophoresis drug delivery system to engage muscarinic receptors with pilocarpine or observed an increase in the slope of internal temperature–sweat rate relation. Pilocarpine LETTERS TO THE EDITOR