Teaching fluid shifts during orthostasis using a classic paper by Foux et al

Hypovolemic and orthostatic challenge can be simulated in humans by the application of lower body negative pressure (LBNP), because this perturbation leads to peripheral blood pooling and, consequently, central hypovolemia. The classic paper by Foux and colleagues clearly shows the effects of orthostasis simulated by LBNP on fluid shifts and homeostatic mechanisms. The carefully carried out experiments reported in this paper show the interplay between different physiological control systems to ensure blood pressure regulation, failure of which could lead to critical decreases in cerebral blood flow and syncope. Here, a teaching seminar for graduate students is described that is designed in the context of this paper and aimed at allowing students to learn how Foux and colleagues have advanced this field by addressing important aspects of blood regulation. This seminar is also designed to put their research into perspective by including important components of LBNP testing and protocols developed in subsequent research in the field. Learning about comprehensive protocols and carefully controlled studies can reduce confounding variables and allow for an optimal analysis and elucidation of the physiological responses that are being investigated. Finally, in collaboration with researchers in mathematical modeling, in the future, we will incorporate the concepts of applicable mathematical models into our curriculum.     

Arterial-alveolar CO2 equilibration in exercising dogs during prolonged rebreathing

Arterial-alveolar equilibration of CO2 during exercise was studied by normoxic CO2 rebreathing in six dogs prepared with a chronic tracheostomy and exteriorized carotid loop and trained to run on a treadmill. In 153 simultaneous measurements of PCO2 in arterial blood (PaCO2) and end-tidal gas (PE'CO2) obtained in 46 rebreathing periods at three levels of mild-to-moderate steady-state exercise, the mean PCO2 difference (PaCO2-PE'CO2) was -1.0 +/- 1.0 (SD) Torr and was not related to O2 uptake or to the level of PaCO2 (30-68 Torr). The small negative PaCO2-PE'CO2 is attributed to the lung-to-carotid artery transit time delay which must be taken into account when both PaCO2 and PE'CO2 are continuously rising during rebreathing (average rate 0.22 Torr/s). Assuming that blood-gas equilibrium for CO2 was complete, a lung-to-carotid artery circulation time of 4.6 s accounts for the observed uncorrected PaCO2-PE'CO2 of -1.0 Torr. The results are interpreted to indicate that in rebreathing equilibrium PCO2 in arterial blood and alveolar gas are essentially identical. This conclusion is at variance with previous studies in exercising humans during rebreathing but is in full agreement with our recent findings in resting dogs.     

Utilization of an Escherichia coli mutant for carbon-13 enrichment of tRNA for NMR studies.

The enrichment of tRNA at specific sites with carbon-13 has been accomplished in vivo using a mutant of Escherichia coli. A relaxed strain of E. coli auxotrophic for methionine was grown in a specifically defined medium supplemented with either [14C] or [13C]-methyl labeled methionine. Cells were collected at the end of the log-phase of growth and tRNA was extracted. Analysis of the radioactivity of the [14C]-labeled tRNA established an incorporation ratio of three labeled carbons per tRNA molecule. Incorporation of the [14C]-label in vivo was confined to the methylation of nucleotides as determined by thin layer chromatography of nucleotides resulting from a ribonuclease digestion of [14C]-labeled tRNA. The carbon-13 NMR spectrum of [13C]-enriched tRNA indicated a similar degree of incorporation into the methylated nucleotides by the substantial enhancement of [13C]-methyl NMR signals only. Assignment of signals has been made for the methyl groups of ribothymidine and N7-methylguanosine in E. coli tRNA.     

Fluctuations in O2 stores and gas exchange with passive changes in posture.

To clarify the role of O2 stores in the fluctuations in VO2 observed with changing posture, O2 intake (Veo2) and pulmonary capillary O2 transfer (Vpco2) were calculated breath by breath with a box-balloon sprometer and mass spectrometer. Changes in O2 stores of the lungs (O2L) and blood (O2b) were computed assuming metabolic rate (Vco2) constant (O2L = Veo2 - Vpco2; O2b = Vpco2 - Vco2). Measurements were made before, during, and after passive tilt to 60 degrees and on return to recumbency after 10 min erect. From supine to upright O2L increased rapidly and O2b dropped slowly, creating a net deficit in Veo2 of 130 ml in 10 min. Return to supine caused rapid loss in O2L and gain in O2b with a net Veo2 excess of 117 ml. Shifts in O2b were 2.5 times greater but opposite to shifts in O2L. Changes in O2b result from shifts in blood volume and flow more than from changes in cardiac output. Refilling of O2b, matching loss while upright, caused transient hypoxia with significant hyperpnea.     

Otoliths of sub-adult Lake Sturgeon Acipenser fulvescens contain aragonite and vaterite calcium carbonate polymorphs

In this study we quantified the percent CaCO₃ polymorph composition in otoliths of larval and juvenile Lake Sturgeon Acipenser fulvescens via X-ray microdiffraction. Sagittal otoliths of sub-adults were primarily composed of aragonite (> 90%) while the lapilli otoliths were 100% vaterite. This is the first time the presence of aragonite in otoliths has been reported in an acipenseriform and is surprising given that the ability to form aragonite otoliths was not thought to have evolved until the separation of teleost and holostean species from other Actinopterygian fishes (e.g., sturgeon, paddlefish, gar).     

Effect of head-down tilt on brain water distribution

Vascular and tissue fluid dynamics in the microgravity of space environments is commonly simulated by head-down tilt (HDT). Previous reports have indicated that intracranial pressure and extracranial vascular pressures increase during acute HDT and may cause cerebral edema. Tissue water changes within the cranium are detectable by T2 magnetic resonance imaging. We obtained T2 images of sagittal slices from five subjects while they were supine and during -13 degrees HDT using a 1.5-Tesla whole-body magnet. The analysis of difference images demonstrated that HDT leads to a 21% reduction of T2 in the subarachnoid cerebrospinal fluid (CSF) compartment and a 11% reduction in the eyes, which implies a reduction of water content; no increase in T2 was observed in other brain regions that have been associated with cerebral edema. These findings suggest that water leaves the CSF and ocular compartments by exudation as a result of increased transmural pressure causing water to leave the cranium via the spinal CSF compartment or the venous circulation.     

Early fluid retention and severe acute mountain sickness.

Field studies of acute mountain sickness (AMS) usually include variations in exercise, diet, and environmental conditions over days and development of clinically apparent edemas. The purpose of this study was to clarify fluid status in persons developing AMS vs. those remaining without symptoms during simulated altitude with controlled fluid intake, diet, temperature, and without exercise. Ninety-nine exposures of 51 men and women to reduced barometric pressure (426 mmHg = 16,000 ft. = 4,880 m) were carried out for 8-12 h. AMS was evaluated by Lake Louise (LL) and AMS-C scores near the end of exposure. Serial measurements included fluid balance, electrolyte excretions, and plasma concentrations, regulating hormones, and free water clearance. Comparison between 16 subjects with the lowest AMS scores near the end of exposure ("non-AMS": mean LL = 1.0, range = 0-2.5) and 16 others with the highest AMS scores ("AMS": mean LL = 7.4, range = 5-11) demonstrated significant fluid retention in AMS beginning within the first 3 h, resulting from reduced urine flow. Plasma Na+ decreased significantly after 6 h, indicating dilution throughout the total body water. Excretion of Na+ and K+ trended downward with time in both groups, being lower in AMS after 6 h, and the urine Na+-to-K+ ratio was significantly higher for AMS after 6 h. Renal compensation for respiratory alkalosis, plasma renin activity, aldosterone, and atrial natriuretic peptide were not different between groups, with the latter tending to rise and aldosterone falling with time of exposure. Antidiuretic hormone fell in non-AMS and rose in AMS within 90 min of exposure and continued to rise in AMS, closely associated with severity of symptoms and fluid retention.