Methylene blue induced O2 consumption is not dependent on mitochondrial oxidative phosphorylation: Implications for salvage pathways during acute mitochondrial poisoning.

While administration of the cyclic redox agent methylene blue (MB) during intoxication by mitochondrial poisons (cyanide, hydrogen sulfide, rotenone) increases survival, the mechanisms behind these antidotal properties remain poorly understood. The objective of the studies presented in this paper was to characterize the interactions between the redox properties of MB, the intermediate metabolism and the mitochondrial respiration. We first show that intra-venous administration of micromolar levels of methylene blue in sedated and mechanically ventilated rats, increases not only resting oxygen consumption but also CO2 production (by ~ 50%), with no change in their ratio. This hypermetabolic state could be reproduced in a cellular model, where we found that the rate of electron transfer to MB was of the same order of magnitude as that of normal cellular metabolism. Notably, the large increase in cellular oxygen consumption caused by MB was relatively indifferent to the status of the mitochondrial respiratory chain: oxygen consumption persisted even when the respiratory chain was inhibited or absent (using inhibitors and cells deficient in mitochondrial oxidative phosphorylation); yet MB did not impede mitochondrial ATP production in control conditions. We present evidence that after being reduced into leuco-methylene blue (LMB) in presence of reducing molecules that are physiologically found in cells (such as NADH), the re-oxidation of LMB by oxygen can account for the increased oxygen consumption observed in vivo. In conditions of acute mitochondrial dysfunction, these MB redox cycling properties allow the rescue of the glycolysis activity and Krebs cycle through an alternate route of oxidation of NADH (or other potential reduced molecules), which accumulation would have otherwise exerted negative feedback on these metabolic pathways. Our most intriguing finding is that re-oxidization of MB by oxygen ultimately results in an in vivo matching between the increase in the rate of O2 consumed, by MB re-oxidation, and the rate of CO2, produced by the intermediate metabolism, imitating the fundamental coupling between the glycolysis/Krebs cycle and the mitochondrial respiration.

[Severe breathlessness in a patient presenting with chronic bronchitis]. / Dyspnée sévère chez un patient présentant une bronchite chronique.

INTRODUCTION: Dyspnoea is the major symptomatic complaint of patients with chronic obstructive pulmonary disease (COPD). Awareness of an unpleasant respiratory sensation, occurs through a complex system of ventilatory control where, as in pain perception, supra-bulbar structures play a major role. In the hyperventilation syndrome, dramatic episodes of dyspnea can occur without any change in peripheral afferent signals and thus do not predictably represent a real deterioration in pulmonary function in patients who also have COPD. CASE REPORT: We report the observation of a patient with GOLD stage 0 COPD who demonstrated a major limitation of daily activities due to supra-bulbar influences on respiratory control which triggered repetitive periods of hyperventilation and generated almost constant dyspnea. CONCLUSIONS: Hyperventilation syndrome can be difficult to diagnose in patients with respiratory disease, although, in most cases, the discrepancy between the severity of symptomatic complaint compared to the degree of alteration of lung function in combination with specific tests may suggest the diagnosis.

Effects of infusion of human methemoglobin solution following hydrogen sulfide poisoning.

RATIONALE: We have recently reported that infusion of a solution containing methemoglobin (MetHb) during exposure to hydrogen sulfide results in a rapid and large decrease in the concentration of the pool of soluble/diffusible H2S in the blood. However, since the pool of dissolved H2S disappears very quickly after H2S exposure, it is unclear if the ability of MetHb to "trap" sulfide in the blood has any clinical interest and relevance in the treatment of sulfide poisoning. METHODS: In anesthetized rats, repetition of short bouts of high level of H2S infusions was applied to allow the rapid development of an oxygen deficit. A solution containing MetHb (600 mg/kg) or its vehicle was administered 1 min and a half after the end of H2S intoxication. RESULTS: The injection of MetHb solution increased methemoglobinemia to about 6%, almost instantly, but was unable to affect the blood concentration of soluble H2S, which had already vanished at the time of infusion, or to increase combined H2S. In addition, H2S-induced O2 deficit and lactate production as well as the recovery of carotid blood flow and blood pressure were similar in treated and control animals. CONCLUSION: Our results do not support the view that administration of MetHb or drugs-induced methemoglobinemia during the recovery phase following severe H2S intoxication in sedated rats can restore cellular oxidative metabolism, as the pool of diffusible sulfide, accessible to MetHb, disappears rapidly from the blood after H2S exposure.

Mechanisms of ventilatory inhibition by exogenous dopamine in cats.

Intravenous injection of dopamine (DA) has consistently been shown to depress minute ventilation (VE). Whereas at low dosage (/=50 microgram/kg). The purpose of this study was to elucidate the mechanisms of DA-induced VE depression. The effects of intravenous injection of DA doses ranging from 1 to 200 microgram/kg were studied in 18 anesthetized cats. DA was injected during air and O2 breathing, after alpha-adrenergic blockade by phenoxybenzamine and after baro- and chemodenervation. VE and CSNCD were also simultaneously recorded on four occasions. In contrast to that with use of low-dose DA, VE depression induced by high-dose DA was dissociated from CSNCD, persisted during 100% O2 breathing, and was significantly correlated with the rise in arterial blood pressure. Although blunted, VE depression was still present after complete chemo- and barodenervation but was suppressed by blocking of the concomitant vasoconstriction with phenoxybenzamine. It is concluded that reflexes of circulatory origin contribute to the VE depression induced by large-dose DA, in addition to its effects on arterial chemoreceptors. The contribution of baroreceptor stimulation and peripheral vasoconstriction is discussed.

O2 uptake kinetics above and below the lactic acidosis threshold during sinusoidal exercise.

O2 uptake (VO2) kinetics at the onset of a constant work rate exercise are difficult to describe for work rates above the lactic acidosis threshold (LAT), because the steady-state level of VO2 response can usually not be identified. To describe the ability of the O2 transport system to deliver and the cells to utilize O2 above the LAT relative to that below the LAT, we applied a fluctuating (sinusoidal) variation of work rate. After 4 min of constant work at the midpoint of the sinusoidal work rate, a fluctuating work rate, at a period of 4 min, was applied below the LAT for the next 16 min. This was repeated in a range of work rates above the LAT with the same sine-wave amplitude. VO2 response appeared to follow a sinusoidal pattern similar to that of work rate for below- and above-LAT exercise. However, the amplitude of the VO2 response was significantly reduced (5.4 +/- 2.6 vs. 7.6 +/- 1.9 ml.min-1 x W-1, P < 0.01), and the phase lag increased above- compared with below-LAT work rate. VO2/heart rate fluctuations were dramatically reduced, whereas heart rate amplitude decreased and phase lag increased, for above-LAT sinusoidal work rate changes. These results suggest that VO2 kinetics are slowed in the work rate domain above the LAT relative to that below the LAT and that VO2 kinetics could be limited by the O2 transport mechanisms to the exercising muscle.

Responses of group III and IV muscle afferents to distension of the peripheral vascular bed.

This study was undertaken to test the hypothesis that group III and IV afferents with endings in skeletal muscle signal the distension of the peripheral vascular network. The responses of these slowly conducting afferents to pharmacologically induced vasodilation and to acute obstruction of the venous drainage of the hindlimbs were studied in barbiturate-anesthetized cats. Afferent impulses arising from endings in the triceps surae muscles were recorded from the L(7) and S(1) dorsal roots. Fifteen of the 48 group IV and 3 of the 19 group III afferents tested were stimulated by intra-aortic injections of papaverine (2-2.5 mg/kg). Sixty-two percent of the afferents that responded to papaverine also responded to isoproterenol (50 microg/kg). Seven of the 36 group IV and 2 of the 12 group III afferents tested were excited by acute distension of the hindlimb venous system. Four of the seven group IV afferents responding to venous distension also responded to papaverine (57 vs. 13% for the nonresponding). Finally, we observed that most of the group IV afferents that were excited by dynamic contractions of the triceps surae muscles also responded either to venous distension or to vasodilatory agents. These results are consistent with the histological findings that a large number of group IV endings have their receptive fields close to the venules and suggest that they can be stimulated by the deformation of these vascular structures when peripheral conductance increases. Moreover, such a mechanism offers the possibility of encoding both the effects of muscle contraction through intramuscular pressure changes and the distension of the venular system, thereby monitoring the activity of the veno-muscular pump.

The V(O2) slow component for severe exercise depends on type of exercise and is not correlated with time to fatigue.

The purpose of this study was to examine the influence of the type of exercise (running vs. cycling) on the O2 uptake V(O2) slow component. Ten triathletes performed exhaustive exercise on a treadmill and on a cycloergometer at a work rate corresponding to 90% of maximal VO2 (90% work rate maximal V(O2)). The duration of the tests before exhaustion was superimposable for both type of exercises (10 min 37 s +/- 4 min 11 s vs. 10 min 54 s +/- 4 min 47 s for running and cycling, respectively). The V(O2) slow component (difference between V(O2) at the last minute and minute 3 of exercise) was significantly lower during running compared with cycling (20.9 +/- 2 vs. 268.8 +/- 24 ml/min). Consequently, there was no relationship between the magnitude of the V(O2) slow component and the time to fatigue. Finally, because blood lactate levels at the end of the tests were similar for both running (7.2 +/- 1.9 mmol/l) and cycling (7.3 +/- 2.4 mmol/l), there was a clear dissociation between blood lactate and the V(O2) slow component during running. These data demonstrate that 1) the V(O2) slow component depends on the type of exercise in a group of triathletes and 2) the time to fatigue is independent of the magnitude of the V(O2) slow component and blood lactate concentration. It is speculated that the difference in muscular contraction regimen between running and cycling could account for the difference in the V(O2) slow component.

Mechanical properties of the upper airway wall in children and their influence on respiratory impedance measurements.

The upper airway wall impedance (Zuaw) may be responsible for a large artifact in the measurement of respiratory system impedance (Zrs) in children. In 17 normal children aged 3.5-13 years Zuaw and Zrs were estimated by varying transrespiratory pressure directly at the mouth (conventional method: Z1) and around the subject's head (head generator method: Z2) from 4 to 32 Hz. Zrs and Zuaw were calculated from Z1 = Zrs.Zuaw/(Zrs+Zuaw) and Z2 = Zrs (1 + Zp/Zuaw), where Zp is the impedance of the pneumotachograph. From the real and imaginary part of Z1, Z2, Zrs, and Zuaw, the corresponding resistance, inertance, compliance and resonant frequency were calculated assuming simple RIC models. No significant difference was found between the mean +/- SE of parameters derived from Zrs (respectively, 6.8 +/- 0.4 cmH2O.L-1.s, 0.034 +/- 0.001 cmH2O.L-1.s2, 10.4 +/- 0.8 m.cmH2O-1, 9.1 +/- 0.3 Hz) and Z2 (6.8 +/- 0.4 cmH2O.L-1.s, 0.038 +/- 0.002 cmH2O.L-1.s2, 10.7 +/- 0.7 ml.cmH2O-1, 8.7 +/- 0.4 Hz). All but the compliance, derived from Z1 were significantly different (P less than 0.01) from those derived from Zrs (5.3 +/- 0.3 cmH2O.L-1.s, 0.008 +/- 0.001 cmH2O.L-1.s2, 11.9 +/- 1.2 ml.cmH2O-1, and 20.3 +/- 1.6 Hz). Respiratory resistance and compliance correlated significantly with height (r = -0.56 and 0.86, respectively), in contrast to upper airway wall resistance (Ruaw) and compliance (Cuaw). Ruaw (8.6 +/- 0.8 cmH2O.L-1.s), Cuaw (1.2 +/- 0.2 m.cmH2O-1), and upper airway wall inertance (0.030 +/- 0.004 cmH2O.L-1.s2) were close to those obtained by direct measurements in adults. The mechanical properties of the upper airway wall are responsible for a significant error in the measurement of Zrs by the conventional method in normal children. Most of the artifact may be corrected for by applying pressure around the child's head.

Heart rate dynamics during sinusoidal exercise: comparison of the control system between children and adults.

The aim was to model the dynamics of heart rate (HR) response to sinusoidal work rate (WR) forcing in children and adults. Seven pre-pubertal boys (aged 10-13) and five adult males (aged 22-37) were studied. Continuous ECG recordings were obtained during the following physiological manoeuvres: five constant amplitude ergometer exercises with WR varying sinusoidally with periods of 0.75, 1, 2, 3.5, and 5 min duration, and one step exercise at a constant WR equal to the midpoint of the sinusoid amplitude. The amplitude ratio (AR; standardized by WR) of the fundamental harmonic of the HR response and the phase shift (phi) between the WR to HR were calculated by Fourier analysis. The HR dynamic parameters (gain and time constant (tau)) of a first order model with or without delay (Td) were also estimated. The AR in children was always higher than that in adults, in absolute terms, but not as a function of body weight. The phi was more delayed in the children than the adults only for the shortest period, i.e. 0.75 min. The tau for the first order model, either without or with Td, was found to be no difference between children and adults (44.7 vs. 45.9 s (without Td), 34.9 vs. 42.3 s (with Td)). Td, however, was longer in the children (6.6 vs. 2.3 s). The goodness of fit for the first order model with Td was better than that without Td in children, i.e. due to the difference of phi for 0.75 min period, whereas the HR dynamics in adults was appropriately described by first order model without Td. It is concluded that the fundamental control of HR to sinusoidal exercise between children and adults was not appreciably different, except for a small Td difference at high sinusoidal frequency.

[Value of pulmonary gas exchange study during exercise in the diagnosis of a muscular glycogenosis]. / Apport de l'étude des échanges gazeux respiratoires au cours de l'exercice dans le diagnostic d'une glycogénose musculaire.

BACKGROUND: The diagnosis of muscular glycogen storage disease is usually difficult to demonstrate as symptoms normally consist of muscular cramps and exercise intolerance. Informations obtained from the study of the pulmonary gas exchange during exercise in a young patient with a glycogen storage disease are reported. CASE REPORT: The ventilatory and gas exchange responses to a cyclo-ergometer exercise were studied in a 17-year-old girl during a ramp-like test (5 W/min). The temporal profile of CO2 production (VCO2) response was clearly abnormal: VCO2 was always lower than oxygen consumption throughout the test, reflecting the lack of lactate buffering by the bicarbonates due to the absence of lactate production. The respiratory ratio was still around 0.75 at the peak of the test. In contrast, responses were perfectly normal in the other members of the family, allowing rejection of the diagnosis of glycogen storage disease without any blood sampling. CONCLUSION: This case illustrates the benefit of studying pulmonary gas exchange during exercise for a non-invasive diagnosis of muscular glycogen storage disease and detection of the disease in the other members of family.

Hypocapnia-dependent facilitation of augmented breaths: observations in awake vs. anesthetized rats.

We investigated whether commonly used injectable laboratory anesthetics alter the regulation of augmented breaths (ABs) in different respiratory backgrounds. Male rats were studied on three separate experimental days, receiving one of three injections in randomized order: ethyl carbamate ('urethane'; 1.2mgkg(-1)), ketamine/xylazine (ket/xyl; 80/10mgkg(-1)), or normal saline. Following each of the three interventions, breathing was monitored during 15min exposures to normoxia (room air), hypoxia (10% O(2)) and hypoxia+CO(2) (10% O(2), 5% CO(2)). Urethane anesthesia completely eliminated ABs from the breathing rhythm in room air conditions (p<0.001), and decreased the hypocapnia-dependent component of this response (p<0.001). ket/xyl left the normal incidence of ABs in room air breathing intact but significantly suppressed the hypoxia-induced facilitation of ABs (p=0.0015). These results provide the first clear evidence that laboratory anesthesia can profoundly alter the regulation of ABs including the hypocapnia-dependent component of their facilitation.

Control of breathing and muscle perfusion in humans.

Very brief and intense exercise triggers a biphasic metabolic and respiratory response with a second phase that occurs after the cessation of the muscular activity. The effects on minute ventilation (V(E)) produced by manipulation of the peripheral circulation in metabolically active muscles could thus be studied without the confounding effects of painful contractions. The second phase of breath-by-breath V(E) and pulmonary gas exchange responses to a brief change in work rate (400 W for 12 s) were studied in six healthy male subjects on four occasions (24 tests). An upper thigh cuff inflation was randomly applied either above or below the systolic blood pressure (200 or 90 Torr, respectively) for 90 s just after the cessation of the contractions prior to the delayed rise in pulmonary gas exchange (eight tests in each subject). Total occlusion produced a significant reduction in the delayed rise in V(E) (-29 +/- 3 %) which normally occurred 20-25 s after the cessation of the contractions. In contrast, cuff inflation at a level predominantly impeding venous return while partially maintaining the arterial supply reduced the rise in pulmonary gas exchange in similar proportion to that during total obstruction but with a slight but not significant reduction in ventilation (-9 +/- 5 %). V(E) during partial occlusion was if anything higher than in control tests with similar oxygen uptake (280 W), despite a higher blood pressure (BP) during occlusion (+7 Torr). It is concluded that the factors resulting from a reduction in venous return or from the involvement of the arterial baroreflex are not responsible for the changes in V(E) produced by the obstruction of the circulation to and from metabolically active muscles. It is proposed that factors related to the level of the perfusion pressure in hyperaemic muscles, possibly located at the venular end of the microcirculation, could account for the changes in V(E) observed.

Improved recovery from laryngeal-stimulation-induced apnea by alpha-adrenergic blockade in adult rabbits.

The apnea triggered by laryngeal stimulation (LS) is associated with a redistribution of blood flow towards the heart and the brain that depends upon peripheral vasoconstriction. The circulatory component of the response to LS has been shown to be reduced following injection of large dose of beta-adrenergic agonist that more intriguingly blunted the ventilatory response as well. To test whether the reduction of LS-induced apnea could be related to the decreased magnitude of the peripheral vasoconstruction, the effects of the alpha-adrenergic antagonist urapidil on the ventilatory and blood pressure responses to LS were studied in 6 adult rabbits. alpha-Adrenergic blockade reduced both the rise in blood pressure and the magnitude of hypopnea induced by LS. These results suggest that the circulatory component of the laryngeal chemoreflex, i.e. the peripheral vasoconstruction, contributes to the magnitude of the ventilatory depression.

Persistent increased lung response to methacholine after normobaric hyperoxia in rabbits.

This study was performed to determine the occurrence and time course of airway hyperreactivity following exposure to normobaric hyperoxia. Twenty-six rabbits were studied. Twelve served as control (group 1), and 14 were exposed to normobaric hyperoxia (FiO2 > or = 95%) for 48 h: 9 rabbits (group 2) were studied after 1 day of recovery in room air and 5 (group 3) after 7 days. The rabbits were anesthetized, curarized and artificially ventilated. Respiratory resistance (Rrs) and elastance (Ers) and their respective changes induced by cumulative doses of aerosolized methacholine were assessed by the multiple linear regression analysis of airway pressure, tidal flow and volume. Weight-specific Rrs and Ers were significantly higher in group 2 (respectively, 87.7 +/- 6.5 cmH2O.L-1.sec.kg and 1100.2 +/- 87.1 cmH2O.L-1.kg, mean +/- SEM) than in group 1 (respectively, 65.2 +/- 3.2 cmH2O.L-1.sec.kg and 904.4 +/- 49.7 cmH2O.L-1.kg (P < 0.05)), but were not different from group 3 (79.4 +/- 7.9 cmH2O.L-1.sec.kg and 952.3 +/- 125.0 cmH2O.L-1.kg, respectively). The dose of methacholine required to increase Rrs by 50% (PDRrs50) was significantly lower in both treated groups: 0.37 +/- 0.11 mg in group 2 and 0.51 +/- 0.19 mg in group 3 vs 2.07 +/- 0.51 mg in group 1 (P < 0.05)). PDErs50 was significantly lower in group 2 (0.45 +/- 0.15 mg) and 3 (0.75 +/- 0.43 mg) compared with controls (1.11 +/- 0.26 mg (P < 0.05)). These results show that hyperoxia induces an increase in Rrs and Ers, and airway hyperreactivity in the rabbit. The latter is prolonged beyond the immediate post-exposure period.

Ventilatory and gas exchange response during walking in severe peripheral vascular disease.

It has long been recognized that at the onset of a dynamic muscular exercise the ventilatory and the circulatory (blood flow) responses appear to be matched, thereby maintaining arterial blood gas homeostasis. Such a coupling has recently been suggested to rely upon ventilatory reflex triggered by mechanoreceptors encoding changes in muscle blood flow or, more likely, blood volume. The aim of this study was to investigate whether patients with severe peripheral blood flow limitation to the lower extremities have a normal ventilatory response during a light intensity exercise. The ventilatory and gas exchange temporal response characteristics were studied during a 6 min walking test in seven patients with severe ischemic peripheral vascular disease and in six normal age-matched subjects. The magnitude of the overall ventilatory and Vo2 increment at the end of the tests was similar in both groups. However, in contrast to the control subjects, who presented an almost rectangular response, the patients had a considerably slowed response dynamics (t50 = 33 +/- 4 vs. 9 +/- 3 sec for Vo2 and 37 +/- 5 vs. 10 +/- 8 sec for VE) with a dramatic reduction in the magnitude of the initial 20 sec of the responses. Although the slow Vo2 dynamics in patients presumably reflected the impeded perfusion of the working muscles. the accompanying sluggishness of the V1 course implies that either muscular ischemia actually inhibits ventilatory response to exercise or, more likely, that this response is strongly linked to the magnitude of the hyperemia in the exercising muscles.