Mesenteric lymphatic vessels adapt to mesenteric venous hypertension by becoming weaker pumps.

Lymphangions, the segments of lymphatic vessels between two adjacent lymphatic valves, actively pump lymph. Acute changes in transmural pressure and lymph flow have profound effects on lymphatic pump function in vitro. Chronic changes in pressure and flow in vivo have also been reported to lead to significant changes in lymphangion function. Because changes in pressure and flow are both cause and effect of adaptive processes, characterizing adaptation requires a more fundamental analysis of lymphatic muscle properties. Therefore, the purpose of the present work was to use an intact lymphangion isovolumetric preparation to evaluate changes in mesenteric lymphatic muscle mechanical properties and the intracellular Ca(2+) in response to sustained mesenteric venous hypertension. Bovine mesenteric veins were surgically occluded to create mesenteric venous hypertension. Postnodal mesenteric lymphatic vessels from mesenteric venous hypertension (MVH; n = 6) and sham surgery (Sham; n = 6) animals were isolated and evaluated 3 days after the surgery. Spontaneously contracting MVH vessels generated end-systolic active tension and end-diastolic active tension lower than the Sham vessels. Furthermore, steady-state active tension and intracellular Ca(2+) concentration levels in response to KCl stimulation were also significantly lower in MVH vessels compared with those of the Sham vessels. There was no significant difference in passive tension in lymphatic vessels from the two groups. Taken together, these results suggest that following 3 days of mesenteric venous hypertension, postnodal mesenteric lymphatic vessels adapt to become weaker pumps with decreased cytosolic Ca(2+) concentration.

Hypercholesterolemia abolishes voltage-dependent K+ channel contribution to adenosine-mediated relaxation in porcine coronary arterioles.

Hypercholesterolemic patients display reduced coronary flow reserve in response to adenosine infusion. We previously reported that voltage-dependent K+ (Kv) channels contribute to adenosine-mediated relaxation of coronary arterioles isolated from male miniature swine. For this study, we hypothesized that hypercholesterolemia attenuates Kv channel contribution to adenosine-induced vasodilatation. Pigs were randomly assigned to a control or high fat/high cholesterol diet for 20-24 wk, and then killed. After completion of the experimental treatment, arterioles (approximately 150 microm luminal diameter) were isolated from the left-ventricular free wall near the apical region of the heart, cannulated, and pressurized at 40 mmHg. Adenosine-mediated relaxation was significantly attenuated in both endothelium-intact and -denuded arterioles from hypercholesterolemic compared with control animals. The classic Kv channel blocker, 4-aminopyridine (1 mM), significantly attenuated adenosine-mediated relaxation in arterioles isolated from control but not hypercholesterolemic animals. Furthermore, the nonselective K+ channel blocker, tetraethylammonium (TEA; 1 mM) significantly attenuated adenosine-mediated relaxation in arterioles from control but not hypercholesterolemic animals. In additional experiments, coronary arteriolar smooth muscle cells were isolated, and whole cell Kv currents were measured. Kv currents were significantly reduced (approximately 15%) in smooth muscle cells from hypercholesterolemic compared with control animals. Furthermore, Kv current sensitive to low concentrations of TEA was reduced (approximately 45%) in smooth muscle cells from hypercholesterolemic compared with control animals. Our data indicate that hypercholesterolemia abolishes Kv channel contribution to adenosine-mediated relaxation in coronary arterioles, which may be attributable to a reduced contribution of TEA-sensitive Kv channels in smooth muscle of hypercholesterolemic animals.

Endogenous testosterone increases L-type Ca2+ channel expression in porcine coronary smooth muscle.

Evidence indicates that gender and sex hormonal status influence cardiovascular physiology and pathophysiology. We recently demonstrated increased L-type voltage-gated Ca2+ current (ICa,L) in coronary arterial smooth muscle (CASM) of male compared with female swine. The promoter region of the L-type voltage-gated Ca2+ channel (VGCC) (Cav1.2) gene contains a hormone response element that is activated by testosterone. Thus the purpose of the present study was to determine whether endogenous testosterone regulates CASM ICa,L through regulation of VGCC expression and activity. Sexually mature male and female Yucatan swine (7-8 mo; 35-45 kg) were obtained from the breeder. Males were left intact (IM, n=8), castrated (CM, n=8), or castrated with testosterone replacement (CMT, n=8; 10 mg/day Androgel). Females remained gonad intact (n=8). In right coronary arteries, both Cav1.2 mRNA and protein were greater in IM compared with intact females. Cav1.2 mRNA and protein were reduced in CM compared with IM and restored in CMT. In isolated CASM, both peak and steady-state ICa were reduced in CM compared with IM and restored in CMT. In males, a linear relationship was found between serum testosterone levels and ICa. In vitro, both testosterone and the nonaromatizable androgen, dihydrotestosterone, increased Cav1.2 expression. Furthermore, this effect was blocked by the androgen receptor antagonist cyproterone. We conclude that endogenous testosterone is a primary regulator of Cav1.2 expression and activity in coronary arteries of males.

Hypercholesterolemia inhibits L-type calcium current in coronary macro-, not microcirculation.

Hypercholesterolemia (HC) is a mary risk factor for the development of coronary heart disease. Coronary ion regulation, especially calcium, is thought to be important in coronary heart disease development; however, the influence of high dietary fat and cholesterol on coronary arterial smooth muscle (CASM) ion channels is unknown. The purpose of this study was to determine the effect of diet-induced HC on CASM voltage-gated calcium current (I(Ca)). Male miniature swine were fed a high-fat, high-cholesterol diet (40% kcal fat, 2% wt cholesterol) for 20-24 wk, resulting in elevated serum total and low-density lipoprotein cholesterol. Histochemistry indicated early atherosclerosis in large coronary arteries. CASM were isolated from the right coronary artery (>1.0 mm ID), small arteries ( approximately 200 microm), and large arterioles ( approximately 100 microm). I(Ca) was determined by whole cell voltage clamp. L-type I(Ca) was reduced approximately 30% by HC compared with controls in the right coronary artery (-5.29 +/- 0.42 vs. -7.59 +/- 0.41 pA/pF) but not the microcirculation (small artery, -8.39 +/- 0.80 vs. -10.13 +/- 0.60; arterioles, -10.78 +/- 0.93 vs. -11.31 +/- 0.95 pA/pF). Voltage-dependent activation was unaffected by HC in both the macro- and microcirculation. L-type voltage-gated calcium channel (Ca(v)1.2) mRNA and membrane protein levels were unaffected by HC. Inhibition of I(Ca) by HC was reversed in vitro by the cholesterol scavenger methyl-beta-cyclodextrin and mimicked in control CASM by incubation with the cholesterol donor cholesterol:methyl-beta-cyclodextrin. These data indicate that CASM L-type I(Ca) is decreased in large coronary arteries in early stages of atherosclerosis, whereas I(Ca) in the microcirculation is unaffected. The inhibition of calcium channel activity in CASM of large coronary arteries is likely due to increases in membrane free cholesterol.

Calcium buffering in coronary smooth muscle after chronic occlusion and exercise training.

OBJECTIVE: Exercise promotes "sarcoplasmic reticulum (SR) Ca2+ unloading" in porcine coronary smooth muscle, resulting in decreased agonist-induced Ca2+ release. We studied Ca2+ handling in healthy, non-occluded right coronary artery cells from hearts chronically occluded at the circumflex artery. METHODS: Myoplasmic free Ca2+ (Ca(m)) was assessed with fura-2 in cells from sedentary (n=8) and aerobically exercise-trained (n=6) female Yucatan pigs after 6-month circumflex artery ameroid occlusion (OCC) and in cells from non-occluded, sedentary pigs (SED, n=5). First, Ca influx was induced by 80 mM KCl depolarization (priming step) followed by 5 mM caffeine to elicit maximal Ca2+ release and depletion. The SR was Ca-loaded again by depolarization and then exposed to caffeine after 2- or 11-min recovery to compare SR Ca2+ unloading. RESULTS: Baseline Ca(m), caffeine-induced peak Ca(m), and depolarization-induced maximum Ca(m) were decreased, and depolarization-induced time-to-half-maximum was increased in OCC vs. SED pigs, suggesting a tonic Ca2+ buffering (lowering) effect of occlusion. Exercise did not alter these effects. SR Ca2+ unloading occurred only in SED, as evidenced by decreased caffeine-induced Ca2+ release after 11 min of recovery, and was inhibited by low extracellular Na+. CONCLUSIONS: SR Ca2+ unloading can be demonstrated in coronary smooth muscle from sedentary pigs using a novel SR Ca2+ unloading protocol, and Ca2+ unloading partly depends on Na+-Ca2+ exchange activity. Furthermore, SR Ca2+ unloading in cells from non-occluded right coronary arteries of chronically circumflex-occluded pig hearts was not altered by exercise, perhaps due to enhanced tonic Ca2+ extrusion versus cells from normal, sedentary animals.

Sarcoplasmic reticulum Ca(2+) uptake is impaired in coronary smooth muscle distal to coronary occlusion.

After chronic occlusion, collateral-dependent coronary arteries exhibit alterations in both vasomotor reactivity and associated myoplasmic free Ca(2+) levels that are prevented by chronic exercise training. We tested the hypotheses that coronary occlusion diminishes Ca(2+) uptake by the sarcoplasmic reticulum (SR) and that exercise training would prevent impaired SR Ca(2+) uptake. Ameroid constrictors were surgically placed around the proximal left circumflex (LCx) artery of female swine 8 wk before initiating 16-wk sedentary (pen confined) or exercise-training (treadmill run) protocols. Twenty-four weeks after Ameroid placement, smooth muscles cells were enzymatically dissociated from both the LCx and nonoccluded left anterior descending (LAD) arteries of sedentary and exercise-trained pigs, and myoplasmic free Ca(2+) was studied using fura 2 microfluorometry. After the SR Ca(2+) store was partially depleted with caffeine (5 mM), KCl-induced membrane depolarization produced a significant decrease in the time to half-maximal (t(1/2)) myoplasmic free Ca(2+) accumulation in LCx versus LAD cells of sedentary pigs. Furthermore, inhibition of sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA; 10 microM cyclopiazonic acid) significantly reduced t(1/2) in cells isolated from the LAD but not from the LCx. Exercise training did not prevent the differences in t(1/2) myoplasmic free Ca(2+) accumulation observed between LCx and LAD cells. Occlusion or exercise training did not alter SERCA protein levels. These results support our hypothesis of impaired SR Ca(2+) uptake in coronary smooth muscle cells isolated distal to chronic occlusion. Impaired SR Ca(2+) uptake was independent of SERCA protein levels and was not prevented by exercise training.

Enhanced L-type Ca2+ channel current density in coronary smooth muscle of exercise-trained pigs is compensated to limit myoplasmic free Ca2+ accumulation.

We hypothesized that enhanced voltage-gated Ca2+ channel current (VGCC) density in coronary smooth muscle cells of exercise-trained miniature Yucatan pigs is compensated by other cellular Ca2+ regulatory mechanisms to limit net myoplasmic free Ca2+ accumulation. Whole-cell voltage clamp experiments demonstrated enhanced VGCC density in smooth muscle cells freshly dispersed from coronary arteries of exercise-trained vs. sedentary animals. In separate experiments using fura-2 microfluorometry, we measured depolarization-induced (80 mM KCl) accumulation of myoplasmic free Ba2+ and free Ca2+. Both maximal rate and net accumulation of free Ba2+ in response to membrane depolarization were increased in smooth muscle cells isolated from exercise-trained pigs, consistent with an increased VGCC density. Depolarization also produced an enhanced maximal rate of free Ca2+ accumulation in cells of exercise-trained pigs; however, net accumulation of free Ca2+ was not significantly increased suggesting enhanced Ca2+ influx was compensated to limit net free Ca2+ accumulation. Inhibition of sarco-endoplasmic reticulum Ca2+-transporting ATPase (SERCA; 10 microM cyclopiazonic acid) and/or sarcolemmal Na+-Ca2+ exchange (low extracellular Na+) suggested neither mechanism compensated the enhanced VGCC in cells of exercise-trained animals. Local Ca2+-dependent inactivation of VGCC, assessed by buffering myoplasmic Ca2+ with EGTA in the pipette and using Ca2+ and Ba2+ as charge carriers, was not different between cells of sedentary and exercise-trained animals. Our findings indicate that increased VGCC density is compensated by other cellular Ca2+ regulatory mechanisms to limit net myoplasmic free Ca2+ accumulation in smooth muscle cells of exercise-trained animals. Further, SERCA, Na+-Ca2+ exchange and local Ca2+-dependent inactivation of VGCC do not appear to function as compensatory mechanisms. Additional potential compensatory mechanisms include Ca2+ extrusion via plasma membrane Ca2+-ATPase, mitochondrial uptake, myoplasmic Ca2+-binding proteins and other sources of VGCC inactivation.

Exercise training restores adenosine-induced relaxation in coronary arteries distal to chronic occlusion.

We previously reported that canine collateral-dependent coronary arteries exhibit impaired relaxation to adenosine but not sodium nitroprusside. In contrast, exercise training enhances adenosine sensitivity of normal porcine coronary arteries. These results stimulated the hypothesis that chronic coronary occlusion and exercise training produce differential effects on cAMP- versus cGMP-mediated relaxation. To test this hypothesis, Ameroid occluders were surgically placed around the proximal left circumflex coronary artery (LCx) of female Yucatan miniature swine 8 wk before initiating sedentary or exercise training (treadmill run, 16 wk) protocols. Relaxation to the cAMP-dependent vasodilators adenosine (10(-7) to 10(-3) M) and isoproterenol (3 x 10(-8) to 3 x 10(-5) M) were impaired in collateral-dependent LCx versus nonoccluded left anterior descending (LAD) arterial rings isolated from sedentary but not exercise-trained pigs. Furthermore, adenosine-mediated reductions in simultaneous tension and myoplasmic free Ca(2+) were impaired in LCx versus LAD arteries isolated from sedentary but not exercise-trained pigs. In contrast, relaxation in response to the cAMP-dependent vasodilator forskolin (10(-9) to 10(-5) M) and the cGMP-dependent vasodilator sodium nitroprusside (10(-9) to 10(-4) M) was not different in LCx versus LAD arteries of sedentary or exercise-trained animals. These data suggest that chronic occlusion impairs receptor-dependent, cAMP-mediated relaxation; receptor-independent cAMP- and cGMP-mediated relaxation were unimpaired. Importantly, exercise training restores cAMP-mediated relaxation of collateral-dependent coronary arteries.

Female acceleration tolerance: effects of menstrual state and physical condition.

INTRODUCTION: The literature contains a paucity of information on female tolerance to high sustained acceleration. With women now flying high-performance aircraft, gender-specific factors that may affect female acceleration tolerance have become increasingly important. The purpose of this investigation was to determine how menstrual state and physical condition affect acceleration tolerance. We hypothesized the menstrual cycle would have no effect on acceleration tolerance and that a positive correlation would exist between physical fitness level and tolerance to high sustained acceleration. METHODS: Centrifuge exposures on 8 female subjects consisted of a relaxed gradual-onset run (0.1 G.s-1) to the visual endpoint, a rapid-onset run (6 G.s-1) to +5 GZ for 15 s, and a +4.5 to +7 GZ simulated aerial combat maneuver (SACM) to physical exhaustion. Acceleration tolerance data were collected at onset of menstruation and 1, 2 and 3 weeks following the onset for two complete menstrual cycles. On separate days, body composition, anaerobic power output and peak oxygen uptake were determined. Retrospective data from 10 male subjects who had performed the +4.5 to +7 GZ SACM were analyzed and compared to these data. RESULTS: Analysis of variance revealed no significant difference in relaxed tolerance or SACM duration between the four selected menstrual cycle time points. Time-to-fatigue on the +4.5 to +7 GZ SACM was positively (p < or = 0.05) correlated with absolute fat-free mass (r = 0.87) and anaerobic power production (r = 0.76) in female subjects. However, when these variables were adjusted for total body mass, the significant correlations no longer existed. No correlation was found between SACM duration and absolute (L min-1) nor relative (ml.kg-1.min-1) aerobic fitness. Time-to-fatigue during the SACM was not significantly different between male and female subjects (250 +/- 97 and 246 +/- 149 s, respectively).

Exercise-enhanced preoxygenation increases protection from decompression sickness.

INTRODUCTION: Prevention of decompression sickness (DCS) during exposure to altitude equivalents of 30,000 ft (9144 m) requires extensive denitrogenation. In preparation for extravehicular activity (EVA), present NASA policy is to denitrogenate using a 10.2 psia staged decompression of the entire shuttle for at least 12 h, including 100 min of preoxygenation (breathing 100% oxygen at 14.7 psia prior to decompression), before decompression to the 4.3 psia (30,000 ft; 9144 m) suit pressure. This staged decompression provides the same or better protection from DCS as a 3.5- or 4-h preoxygenation used on earlier Shuttle EVA's. For high altitude reconnaissance flights at similar cockpit altitudes, a 1-h preoxygenation is currently required. METHODS: We have investigated the use of a 1-h and a 15-min preoxygenation period, each beginning with 10 min of dual-cycle ergometry performed at 75% of each subject's peak oxygen consumption (VO2peak) to enhance preoxygenation efficiency by increasing perfusion and ventilation. Male subjects accomplished a 1-h preoxygenation with exercise, a 15-min preoxygenation with exercise, or a 1-h resting preoxygenation before exposure to 4.3 psia for 4 h while performing light to moderate exercise. RESULTS: Incidence of DCS following the 1-h preoxygenation with exercise (42%; n = 26) was significantly less than that following the 1-h resting preoxygenation (77%; n = 26). Incidence and onset of DCS following the 15-min preoxygenation with exercise (64%; n = 22) was not significantly different from the incidence following the 1-h resting control. CONCLUSION: Preoxygenation with exercise has been shown to provide significantly improved DCS protection when compared with resting preoxygenation.

Physiological responses of rhesus monkeys to exercise at varied temperatures.

This research characterizes the effects of selected physiological stressors such as work and various environmental heat loads in rhesus monkeys. Non-human primates (N = 6) were behaviorally conditioned to exercise in a wheel ergometer at approximately 3 METs (1 MET = 3.5 ml O2 uptake.kg-1.min-1). On separate days, each monkey attempted six work/rest cycles (10 min work: 1 min rest) at 15, 25, and 35 degrees C (Tdb), vapor pressure < 10 mm Hg. Core temperature (Tco), body weight (BW) and blood samples were taken immediately before and after exercise. Excessively high heat storage rates dictated that the 35 degrees C trial be limited to three work/rest bouts. The change in Tco during exercise was significantly greater in the 35 degrees C trial than during the 15 and 25 degrees C work bouts. Sweat rate, as determined by the change in BW over time, was also significantly greater during the 35 degrees C trial as compared to the 15 and 25 degrees C trials. Glucose levels (mean +/- SE) declined significantly during exercise from 4.35 +/- 0.1 and 4.58 +/- 0.4 mmol to surprisingly low levels of 1.67 +/- 0.2 and 1.76 +/- 0.2 mmol in the 15 and 25 degrees C trials, respectively. Increases in blood lactate, glycerol, and triglycerides were observed independent of environmental temperature. Free fatty acids increased during exercise in the 15 and 25 degrees C trials but declined slightly during the shorter 35 degrees C trial. The findings of this study indicate that the increased heat storage observed at higher environmental heat loads appears to substantially limit the amount of work these primates can perform, possibly the result of a limited sweat production capacity. Additionally, plasma glucose following exercise decreased to levels not typically seen in humans. The general metabolic profile in these primates was otherwise similar to that observed in humans at this work level.

Hypohydration causes cardiovascular drift without reducing blood volume.

To determine the effect of hydration on cardiovascular drift (i.e.; increased heart rate and reduced stroke volume) during exercise in a 21 degrees C environment, nine subjects were studied while cycling at 65% of peak oxygen consumption when euhydrated and while still hypohydrate, following exercise-induced dehydration and a 2 h rest/rehydration period. Subjects dehydrated by exercising in the heat (32 degrees C) until body weight was reduced 2.5%. On two separate occasions following exercise, subjects either received no fluid or ingested a volume of water equal to 100% of the fluid lost during exercise. Following the 2 h rest/rehydration period, 65 +/- 6% of the ingested water was retained and thus the subjects were hypohydrated by 0.9 +/- 0.1%, compared to being hypohydrated by 2.8 +/- 0.1% when no fluid was ingested. Despite these differences in whole body hydration, blood volume during exercise remained at euhydrated levels when hypohydrated by 0.9% and 2.8%. However, the degree of cardiovascular drift was graded in proportion to hypohydration. Compared to the responses when euhydrated, heart rate was elevated 10 +/- 2 and 18 +/- 2 bt/min, whereas stroke volume was reduced 9 +/- 3 and 18 +/- 2 ml/bt, respectively, when hypohydrated by 0.9% and 2.8% during the water and no fluid trials (p < 0.05 for all comparisons). These observations indicate that cardiovascular drift during exercise in a 21 degrees C environment is graded in proportion to hydration and, under these conditions, not due to reductions in blood volume.

Rehydration after exercise with common beverages and water.

This study assessed the effectiveness of two common rehydration beverages (a caffeinated diet cola (DC) and a 6% carbohydrate-electrolyte (CE) solution) compared with water (W) for whole body rehydration, gastric emptying and blood volume (BV) restoration during a 2 h rehydration period following exercise-induced dehydration. Subjects (mean VO2max = 4.2 +/- 0.6 l.min-1.min-1; n = 19) exercised at 60-80% VO2max in the heat (32 degrees C; 40% rh) until approximately 2.5% (1.95 +/- 0.12 kg) of their body weight (BW) was lost. After exercise, the subjects sat for 2 h in a thermoneutral environment (21 degrees C; 60% rh) and drank a volume of DC, W and CE equal to the fluid lost. Fluids were consumed in two boluses averaging 1,046 +/- 198 and 912 +/- 186 ml at 0 and 45 min of the 2 h rehydration period, respectively. At the end of the rehydration period, no fluid remained in the stomach during any of the trials as indicated by epigastric impedance. However, in all the trials the subjects were somewhat hypohydrated (range 0.6-0.9 kg BW below euhydrated BW; p less than 0.05) after the 2 h rehydration period since additional water and BW were lost as a result of urine formation, respiration, sweat and metabolism. The percentage of body weight loss that was regained (used as an index of % rehydration) during DC (54 +/- 5%) was significantly lower than that of W and CE (64 +/- 5% and 69 +/- 5%, respectively; p less than 0.05; n = 10).(ABSTRACT TRUNCATED AT 250 WORDS)