S. Howell

Effects of neostigmine and salbutamol on diaphragmatic fatigue

We studied the effects of neostigmine and salbutamol on the force generated by the fatigued diaphragm in anesthetized dogs. Mechanically ventilated animals were prepared with an open thorax. A thin-walled latex balloon was positioned beneath the diaphragm to measure transdiaphragmatic pressure (Pdi) and a rigid cast was fixed around the abdomen to limit changes in diaphragmatic length and geometry during contractions. Pdi was the index of force generated by the diaphragm. We measured Pdi during supramaximal phrenic stimulation at different frequencies and during spontaneous inspiratory efforts. The diaphragm was fatigued by repeated phrenic stimulation. Fatigue significantly reduced Pdi at all frequencies of stimulation and during spontaneous contractions (P less than 0.05). The reduction in Pdi was associated with a decrease in peak twitch tension (PTT) to 50% of control (P less than 0.05). Infusion of neostigmine restored PTT to values equivalent with or greater than control (P less than 0.05) and improved Pdi at low stimulation frequencies (P less than 0.05) and during spontaneous inspiratory efforts (P less than 0.05). Infusion of salbutamol had no effect on PTT, but did significantly shortened twitch half relaxation time (P less than 0.05). Salbutamol also had no effect on Pdi during stimulated and spontaneous contractions. We conclude that neostigmine improves force generated by the fatigued diaphragm by increasing twitch amplitude while salbutamol did not have a positive inotropic effect.

Compartmental analysis of steady-state diaphragm Ca2+ kinetics in chronic congestive heart failure.

UNLABELLED An analytic method based on simulation and modeling of long-term 45Ca(2+) efflux data was used to estimate steady-state Ca(2+) contents (nmolCa(2+)g(-1)tissuewetwt.) and exchange fluxes (nmolCa(2+)min(-1)g(-1)tissuewetwt.) for extracellular and intracellular compartments in in vitro resting diaphragm from congestive heart failure (CHF, n=12) and sham-operated (SHAM, n=10) rats. Left hemidiaphragms were excised from experimental animals, loaded with 45Ca(2+) for 1h, and washed out with 45Ca(2+)-free perfusate for 8h. Tissue from the right hemidiaphragm was used to assess single-fiber cross-sectional area (CSA) as well as the relative proteolytic activity of Ca(2+)-dependent calpain. Kinetic analysis of 45Ca(2+) efflux data revealed that CHF was associated with increased Ca(2+) contents of extracellular and intracellular compartments as well as increased Ca(2+) exchange fluxes for all compartments. This accounted for the model prediction of a 250% increase in total diaphragm Ca(2+). Furthermore, single-fiber CSA was decreased 12% and proteolytic activity of calpain was increased twofold in CHF diaphragm relative to SHAM. CONCLUSIONS The kinetic data are consistent with the hypothesis that diaphragm Ca(2+) overload in CHF required all intercompartmental Ca(2+) fluxes to increase. The potential relationships among Ca(2+) overload, increased activity of calpain, and wasting of the diaphragm in CHF are discussed.

Skeletal muscle hemoglobin content measured by near-infrared spectroscopy during oscillatory venous pressure variation

Changes in oxidized (HbO), reduced (Hbr), and total hemoglobin (Hbt) contents were monitored by near infrared spectroscopy in human forearm skeletal muscle during oscillatory variations of the effective venous pressure. Laser diode pulses (wavelength, 775, 810, 865, 904; duration, 60 ns) were directed to the muscle by means of an optic fiber bundle and detected with a sensing fiber positioned at 1.5 cm from the emitting bundle. Sinusoidal pressure waves (frequency, 1 and 2 cycles/min; amplitude, 10-15 mm Hg) generated by a piston pump were transmitted to a sphygmomanometer cuff placed on the arm, the mean cuff pressure (Pc) being 20, 40 and 60 mm Hg. Variations of HbO, Hbr and Hbt were computed from the optical signals and processed by Fourier analysis to characterize their amplitude and phase relative to the cuff mean pressure oscillations (Posc). Oscillations of HbO, Hbr, Hbt were observed for all mean cuff pressures, the amplitude of the variations being decreased with increasing Pc. For Pc equals 20 mm Hg, the oscillations of HbO and Hbt were nearly in phase with Posc whereas the oscillation of Hbr were out of phase with HbO and Posc. Increasing Pc resulted in an increase of the phase difference between HbO and Posc, Hbr remaining out of phase with Posc. These trends could be predicted with a lumped model of the forearm vasculature, suggesting that the technique could be used to asses mechanical characteristics of vascular beds.

NMR study of rat diaphragm exposed to metabolic and compensated metabolic acidosis

When exposed to hypercapnia, several muscles deteriorate with respect to their mechanical performance. Exposure to metabolic acidosis and, perhaps surprisingly, to compensated metabolic acidosis has the same effect on the diaphragm. The mechanisms involved in these effects remain unclear. If the diaphragmatic intracellular pH (pHi) is assumed to decrease with hypercapnia, to remain unchanged during metabolic acidosis, and to increase during compensated metabolic acidosis, it would appear that different mechanisms must be responsible for the depreciation in the diaphragms mechanical performance. The present experiments using 31P nuclear magnetic resonance (31P-NMR) spectroscopy were undertaken to determine the effect of metabolic acidosis and compensated metabolic acidosis on pHi and on high-energy phosphate metabolites in the resting rat diaphragm. A whole diaphragm was slightly stretched while being stitched onto a fiberglass mesh. The area approximated that at functional residual capacity. It was superfused in the NMR sample tube with a phosphate-free Krebs-Ringer bicarbonate solution [( HCO3-] = 6 meqO equilibrated with either 95% O2-5% CO2 or 98.75% O2-1.25% CO2). Spectra were acquired during 15-min intervals for control (30 min of normal Krebs-Ringer bicarbonate superfusate, equilibrated with 95% O2-5% CO2), for 120 min of exposure to either form of acidosis and for 60 min of recovery with normal superfusate. The pHi decreased rapidly during metabolic acidosis but did not change significantly during compensated metabolic acidosis. In both forms of acidosis, phosphocreatine declined gradually but not significantly, whereas ATP and inorganic phosphate did not change at all. The results suggest that HCO3- passes freely through the diaphragmatic sarcolemma, very much like the cardiac sarcolemma.(ABSTRACT TRUNCATED AT 250 WORDS)