Pulmonary blood flow redistribution by increased gravitational force.

This study was undertaken to assess the influence of gravity on the distribution of pulmonary blood flow (PBF) using increased inertial force as a perturbation. PBF was studied in unanesthetized swine exposed to -Gx (dorsal-to-ventral direction, prone position), where G is the magnitude of the force of gravity at the surface of the Earth, on the Armstrong Laboratory Centrifuge at Brooks Air Force Base. PBF was measured using 15-micron fluorescent microspheres, a method with markedly enhanced spatial resolution. Each animal was exposed randomly to -1, -2, and -3 Gx. Pulmonary vascular pressures, cardiac output, heart rate, arterial blood gases, and PBF distribution were measured at each G level. Heterogeneity of PBF distribution as measured by the coefficient of variation of PBF distribution increased from 0.38 +/- 0.05 to 0.55 +/- 0.11 to 0.72 +/- 0.16 at -1, -2, and -3 Gx, respectively. At -1 Gx, PBF was greatest in the ventral and cranial and lowest in the dorsal and caudal regions of the lung. With increased -Gx, this gradient was augmented in both directions. Extrapolation of these values to 0 G predicts a slight dorsal (nondependent) region dominance of PBF and a coefficient of variation of 0.22 in microgravity. Analysis of variance revealed that a fixed component (vascular structure) accounted for 81% and nonstructure components (including gravity) accounted for the remaining 19% of the PBF variance across the entire experiment (all 3 gravitational levels). The results are inconsistent with the predictions of the zone model.

A conscious baboon model for evaluation of hemodynamics in altered gravity.

A model was developed for evaluation of cardiovascular parameters in conscious baboons exposed to altered gravitational environments. Baboons were trained to sit quietly in a confinement chair of unique design which allowed a range of normal physical activity. They were then instrumented with high-fidelity blood pressure transducers in the aorta and left ventricle, electromagnetic flow probes encircling the proximal ascending aorta, left and right atrial fluid catheters, left ventricular sonomicrometer crystals in a 3-axis configuration, and a hydraulic occluder cuff encircling the inferior vena cava. Catheters and transducer wires were exteriorized at the midscapular region of the back. Viability of percutaneous exit sites was enhanced by use of velour cuffs on the transducer wires, providing a scaffold for wound healing. Pressure transducers and flow probes were calibrated and balanced during postoperative cardiac catheterization procedures. This instrumentation allowed measurement of beat-to-beat stroke volume and cardiac output not reliant on thermodilution techniques. Postoperative longevity was from 1 to 10 months. Instrumentation failure included endocardial trapping of ventricular pressure transducers, corrosion of ventricular sonomicrometer crystals, and catheter tip thrombosis. Acquisition of high quality data was possible with this model in several different environments of altered gravitational stress, allowing characterization of aortic flow and ventricular performance.

An acute animal model that simulates the hemodynamic situations present during +Gz acceleration.

Air combat maneuver acceleration (G) profiles with onset/offset patterns that occur faster than the response characteristics of the human cardiovascular system may lead to regulatory instability and, ultimately, acceleration-induced loss of consciousness (G-LOC) incidents. We have developed an acute animal model that simulates the hemodynamic situations seen under acceleration to study the effects of complex G environments on individual reflexogenic areas. This preparation allowed us to individually isolate the effects of high gravity on venous return and cardiac preload, arterial baroreflexes and splanchnic capacity. This report describes the preparation and presents examples of the types of +Gz simulations possible and recordings of the responses of the animals. Further, we tested the hypothesis that the volume of blood displaced from the cephalic regions of the circulation and the rate of displacement into the splanchnic capacitance with G onset is affected by distending pressure at the carotid/aortic baroreceptor sites. Early results from 7 dogs show that resistance to flow into the splanchnic beds is affected by changes in distending pressure occurring at arterial baroreceptor sites. When pressure distending the carotid/aortic baroreceptors was increased, resistance to flow into the abdominal vascular beds was decreased. This result suggests that sudden increases in +Gz loads occurring during the overshoot phase from a previous G-peak may result in reduced tolerance.

Beat-to-beat determination of peripheral resistance and arterial compliance during +Gz centrifugation.

This study focused on the problem of describing changes in total peripheral resistance (TPR) and systemic arterial compliance (SAC) under time-varying +Gz acceleration stress. Nonsteady-state measures of peripheral resistance can only be derived when arterial compliance is taken into account. We have developed a successful analytical model to track simultaneous changes in peripheral resistance and systemic arterial compliance during non-stationary periods of increased gravitational load on a beat-to-beat basis. Using a 2-element windkessel model, aortic flow into an input node was defined as equal to the sum of a capacitative (Cao) and a resistive (Rarterial) flow leaving the node such that: Iao = Caod(Pao - Ppleural)/dt + (Pao - Pra)/Rarterial We made the assumption that Cao and Rarterial were constant over a cardiac cycle, and divided the pressure and flow signals for each beat of a record into two different intervals, integrating this equation over each, giving two equations in two unknowns. Cao and Rarterial were then obtained from the matrix solutions. To test the model, we used recordings from chronically instrumented baboons subjected to a 10 s rapid onset +Gz (head-to-foot) stress. Beat-to-beat calculations of peripheral resistance and systemic arterial compliance from our model were compared to values obtained from a previously reported 3-element wind-kessel model.

Differences between syncope resulting from rapid onset acceleration and orthostatic stress.

BACKGROUND AND HYPOTHESIS: Orthostatically-induced syncope is accompanied by venous pooling and vasodilatation. Loss of consciousness during head-to-foot acceleration (G-LOC) in aviators may be caused by a different mechanism, as venous pooling should be prevented through the use of an anti-G suit. This research was conducted to test the hypothesis that in individuals wearing a well-fitted anti-G garment, no important changes occur in the volume of dependent regions during loss of consciousness resulting from rapid onset acceleration stress. Further, this work compares venous pooling patterns in G-LOC subjects to patterns seen during syncope in volunteers and patients subjected to orthostatic stress. We conducted the tilt/LBNP tests to establish what level of venous pooling was required to induce syncope in the absence of a hydrostatic component (other than 1 G) and to confirm that our equipment was sensitive enough to detect volume changes large enough to cause syncope. METHODS: Shifts in blood volume to the calf, thigh and abdominal segments were compared in subjects with G-LOC to those in subjects taken to presyncope with orthostatic stress created by upright tilt and lower body negative pressure (LBNP). Centrifuge subjects were exposed to a 15 s rapid onset (6 G.s-1) + 5 Gz exposure on the centrifuge while remaining relaxed and wearing a well-fitting anti-G suit, but with the anti-G suit pressure inactivated. RESULTS: Blood volume decreased an average of 14.9 +/- 22.1 ml in the calf segment; increased an average of 64.1 +/- 7.9 ml in the thigh segment, and decreased an average of 80.1 +/- 29.7 ml in the abdominal segment. The mean net change in volume of the three combined regions was not significantly different from zero. Presyncope was induced in subjects by a progressive exposure to upright tilt, and then addition of LBNP at -20 mm Hg and -40 mm Hg. In the tilt/LBNP group, there was a net increase of 1022 +/- 269.8 ml for the combined segments. Changes in all three segments were significantly different than the mean segmental volume changes seen in centrifuge subjects at G-LOC endpoints. Significant changes from baseline mean arterial pressure, but not heart rate were also seen within, but not between the 2 groups, with mean eye level blood pressures (ELBP) falling an average of 45.6 +/- 7.7 mm Hg in the tilt/LBNP group at syncope and 105.1 +/- 15.5 mm Hg in the centrifuge subjects at G-LOC. CONCLUSIONS: These differences suggest that G-LOC may be due entirely to hydrostatic effects, with venous pooling being prevented by the wearing of an ant-G garment, even when it remains uninflated.

Relaxation-induced contraction of smooth muscle surrounding hamster ovarian follicles.

Changes in intrafollicular pressure and follicular diameter resulting from injecting or withdrawing fluid from the antrum were measured in preovulatory follicles and used as an assay for changes in tension in the follicular wall by applying the Laplace relationship for thin-walled spheres. Passive length-tension curves were constructed from pressure-volume measurements to establish baseline wall stiffness. Any subsequent change in pressure could then be compared to the length-tension curves to evaluate whether it arose from active tension development or from passive stretch. When intact follicles (1-2h before ovulation) were subjected to release of passive stretch, they exhibited a contractile response that lasted 15 sec-2 min and was characterized by cyclic increases and decreases in tension, with a period of 1 cycle every 2-3 sec. The probability of activating a response in the tissue was most strongly correlated with the rate of release of passive stretch. Intrafollicular pressures generated during active contractile responses sometimes reached 80 mmHg (10.64 mPa), corresponding to a wall tension of 5332 dynes/cm (5.332 N/m) (for a 1 mm follicle) and were clearly well above the passive length-tension curves. Passive stretching of the follicular wall during a contractile response to 5-hydroxytryptamine stimulation resulted in large reductions in active wall tension for the duration of the stretch. These results are consistent with a stretch-activated inhibition of contractile events.

Hamster thecal cells express muscle characteristics.

Contraction of the follicular wall about the time of ovulation appears to be a coordinated event; however, the cells that mediate it remain poorly studied. We examined the theca externa cells in the wall of hamster follicles for the presence of a functional actomyosin system, both in developing follicles and in culture. We used a monoclonal antibody (HHF35) that recognizes the alpha and gamma isoelectric variants of actin normally found in muscle, but not the beta variant associated with non-muscle sources, to evaluate large preovulatory follicles for actin content and composition. Antibody staining of sectioned ovaries showed intense circumferential reactivity in the outermost wall of developing follicles. Immunoblots from two-dimensional gels of theca externa lysates demonstrated the presence of the two muscle-specific isozymes of actin. Immunofluorescence of cultured follicular cells pulse-labeled with [3H] thymidine (for autoradiographic detection of DNA replication) revealed the presence, in many dividing cells, of actin filaments aligned primarily along the longitudinal axis of the cells. In cultures exposed to the calcium ionophore A23187 (10(-4) M) for varying periods (5 min to 1 h), contraction of many individual muscle-actin-positive cells was observed. Immunofluorescence of these cells, fixed immediately after ionophore-induced contraction, revealed compaction of the actin filaments. Our findings demonstrate that the cells of the theca externa contain muscle actins from an early stage and that these cells are capable of contraction even while proliferating in subconfluent cultures. They suggest that follicular growth may include a naturally occurring developmental sequence in which a contractile cell type proliferates in the differentiated state.

Slurry sampling for the determination of arsenic, cadmium, and lead in mainstream cigarette smoke condensate by graphite furnace-atomic absorption spectrometry and inductively coupled plasma-mass spectrometry.

The slurry sampling technique has been applied for the determination of As, Cd, and Pb in mainstream cigarette smoke condensate (MS CSC) by graphite furnace-atomic absorption spectrometry (GF-AAS) and inductively coupled plasma-mass spectrometry (ICP-MS). The MS CSC of the 1R4F Reference Cigarette was collected by electrostatic precipitation and was subsequently prepared as two slurry samples with and without the dispersing agent Triton X-100. Comparison of results determined by ICP-MS analyses of the 1R4F MS CSC slurry samples with those from the conventional microwave digestion method revealed good agreement. The precision of Triton X-100 slurry sampling and of microwave-assisted digestion was better than 10% RSD, and both were superior to slurry sampling without use of Triton X-100. The accuracy of the analytical results for the Triton X-100 slurry sample was further verified by graphite furnace-atomic absorption spectrometry (GF-AAS). For GF-AAS, the method limits of detection are 1.6, 0.04, and 0.5 microg x L(-1) for As, Cd, and Pb, respectively. For ICP-MS, the method limits of detection are 0.06, 0.01, and 0.38 microg x L(-1) for As, Cd, and Pb, respectively. The MS CSC of the 1R4F Reference Cigarette was collected in accordance with the Federal Trade Commission (FTC) smoking regime (35 mL puff volume of 2-s puff duration at an interval of 60 s) and the concentrations of As, Cd and Pb were 6.0+/-0.5, 69.3+/-2.8, and 42.0+/-2.1 ng/cigarette, respectively.

Stroke-prone SHR vascular muscle Ca2+ current amplitudes correlate with lethal increases in blood pressure.

Studies on the possible causal relationship between the Ca2+ channel current density in the vascular muscle cell (VMC) and increases in blood pressure were extended by a comparison of stroke-prone spontaneously hypertensive rats (SP-SHR) with N/nih outbred normotensive rats. Maximal amplitudes of both L-type and T-type Ca2+ channel currents were significantly increased in SP-SHR without a difference in cell capacitance. SP-SHR peak current amplitudes in 20 mM Ba2+ averaged 446 +/- 64 pA while N/nih averaged 156 +/- 25 pA (clearly separated statistically). Both L-type and T-type Ba2+ currents (IBa) were significantly increased in SP-SHR, shown also by peak current frequency distributions. There was a significant shift to the left of both activation (7 mV) and inactivation (15 mV) current-voltage (I-V) plots. SP-SHR IBa recovery from inactivation was significantly slower (103 versus 61 ms) than in N/nih VMC. The increases in SP-SHR IBa amplitude under maximized conditions correlated with increases in blood pressure. Together with earlier observations of increased vascular muscle Ca2+ current density coexistent with blood pressure elevation in Kyoto-Wistar SHR, these data provide evidence for altered function of Ca2+ channels as a fundamental component of hypertension. Since the Ca2+ channel alterations exist in venous VMCs of newborn SP-SHR rats (in a low pressure blood vessel and at a time when increased Ca2+ current density could not be an effect of increased blood pressure), our results add to the growing evidence of Ca2+ channel abnormalities as a cause of genetic hypertension.

Reflex vascular responses to alterations in abdominal arterial pressure and flow in anaesthetized dogs.

The existence of abdominal arterial baroreceptors has long been controversial. Previously difficulties have been encountered in localizing a stimulus to abdominal arteries without affecting reflexogenic areas elsewhere. In these experiments, using anaesthetized dogs, the abdomen was vascularly isolated at the level of the diaphragm, perfused through the aorta, and drained from the inferior vena cava to a reservoir. Changes in abdominal arterial pressure were effected by changing the perfusion pump speed. During this procedure the flow back to the animal from the venous outflow reservoir was held constant. Increases and decreases in abdominal arterial pressure resulted, respectively, in decreases and increases in perfusion pressure to a vascularly isolated hind-limb and in some dogs also a forelimb. Responses were significantly larger when carotid sinus pressure was high (120-180 mmHg) than when it was low (60 mmHg). Responses were still obtained after cutting vagus, phrenic and splanchnic nerves, but were abolished by spinal cord lesion at T12. These experiments provide evidence for the existence of abdominal arterial baroreceptors. The afferent pathway for the reflex vasodilatation appears to run in the spinal cord.

Reflex vascular responses to abdominal venous distension in the anesthetized dog.

This was undertaken to determine whether distension of the subdiaphragmatic veins results in reflex vasoconstriction and interacts with the carotid baroreflex. In alpha-chloralose-anesthetized open-chest dogs, a perfusion circuit controlled carotid and thoracic aortic pressures, splanchnic and limb blood flows, and cardiopulmonary blood flows. At carotid sinus pressures below approximately 90 mmHg, increases in splanchnic pressure of 7 mmHg or more resulted in increases in vascular resistance in both the splanchnic and limb circulations; there was no response at higher carotid pressures. At high venous pressures, the average maximum gains of the carotid baroreflex for splanchnic and limb resistance responses were increased by 106 and 67%, respectively. The responses were not abolished by cutting the vagal or phrenic nerves but were prevented by cutting the splanchnic nerves and, for the limb, the sciatic and femoral nerves. These results suggest that splanchnic congestion, by causing vasoconstriction and augmentation of the carotid baroreflex, may be important in the maintenance of blood pressure during gravitational stress.

Effect of upright tilt on ventricular/vascular coupling in chronically instrumented primates.

Studies of the hydraulic loading conditions on the heart in humans, especially pulsatile load, have primarily been limited to the supine state. Therefore, we have chosen a nonhuman primate model, the baboon, to assess left ventricular/vascular coupling in both supine and upright positions. Primate subjects were studied by catheterization under sedation and then after surgical implantation of transducers. This allowed the evaluation of postural stress in the chronically instrumented conscious baboon and then after light dissociative doses of ketamine. Basic hemodynamic variables were evaluated for baboons in supine and upright positions. Fourier analysis was applied to aortic pressure and flow to obtain input and characteristic impedance and the ratio of pulsatile (Wp) to total (Wt) left ventricular power (Wp/Wt). The aortic reflected, or backward, pressure was also calculated. Peripheral resistance increased (P = 0.01) and reflected pressure decreased (17.74 +/- 1.50 vs. 15.21 +/- 2 mmHg; P < 0.01) in upright subjects. Characteristic impedance and Wp/Wt were unchanged. Postoperatively, peripheral resistance increased (2,651 +/- 311 vs. 3,667 +/- 276; P < 0.05) and mean power and Wt decreased (P < 0.01) without changes in reflected pressure. All variables were unchanged after light dissociative doses of ketamine. Thus there is no significant change in efficiency of left ventricular/vascular coupling formulated in terms of Wp/Wt or input impedance with postural stress.

Reflex vascular responses in the anesthetized dog to large rapid changes in carotid sinus pressure.

This study examined reflex vascular responses to large rapid increases and decreases in carotid sinus pressure to determine whether delayed or inappropriate vascular responses might be obtained that, if they occurred in people, could lead to hypotension during exposure to rapidly alternating gravitational forces. In chloralose-anesthetized open-chest dogs, a perfusion circuit controlled carotid sinus and thoracic aortic pressures and blood flows to both the vascularly isolated abdominal circulation and a hindlimb (perfusion pressure changes denoted resistance). When carotid pressure was increased and decreased over the range of 60-180 mmHg, the resulting reflex vasodilatation occurred significantly more rapidly than the vasoconstriction (P < 0.001). In the abdominal vascular bed, time constants for vasodilatation and vasoconstriction were 4.2 +/- 0.5 and 7.5 +/- 1.0 s, respectively. Decreases in carotid pressure in pulses of 10-s duration or less failed to elicit maximal vasoconstriction, whereas increases in carotid pressure lasting as little as 5 s did elicit maximal vasodilatation. "Square-wave" alternations in carotid pressure with periods of 10 s or less (5 s high, 5 s low) resulted in attenuation of the vasoconstriction, and at a 4-s period, both vascular beds remained almost maximally vasodilated throughout. The failure of vascular resistance to follow carotid pressure changes was not due to a failure of the response of sympathetic efferent activity, since the time constants for the reduction and increase in discharge were much shorter at 0.56 +/- 0.13 and 0.43 +/- 0.10 s, respectively. These results indicate that rapid changes in carotid pressure could result in inappropriate vasodilatation and hypotension and might, in some circumstances, such as in pilots flying high-performance aircraft, predispose to syncope.