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.

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.

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.

[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.

Inhibition of midbrain-evoked tonic and rhythmic motor activity by cutaneous stimulation in decerebrate cats.

The effect of mechanical and electrical stimulation of cervical cutaneous afferents was analysed on both the centrally induced tonic and rhythmic activities in hindlimb antagonist muscle nerves of 16 decerebrate paralysed cats. Electrical stimulation of dorsal midbrain evoked in the nerve to the tibialis anterior muscle (TAn) either rhythmic discharges (n=14), associated with tonic discharges in ten cats, or only tonic discharges (n=4). Centrally induced activity in the ipsilateral nerve to gastrocnemius medialis (GMn) occurred in fewer cats (n=12) and displayed similar patterns as in TAn. Manual traction of the scruff of the neck reduced the TAn tonic and rhythmic discharges (n=6) by 73% (P<0.05) and 71% (P<0.05), respectively, and reduced only the tonic component of GMn discharges (by 41%, n=3). Electrical stimulation (impulses 0.1-0.5 ms, 50 Hz) of cervical nerves belonging to C5 or C6 dermatomes, the intensity (0.4-4 mA) of which induced minimal inhibition of both TAn and GMn discharges, reduced significantly the tonic component of TAn discharges (by 39%, n=4). At higher intensities of electrical cervical nerve stimulation (2-6 mA) inducing maximal inhibitory effect, both tonic and rhythmic activities in TAn and GMn were both significantly reduced by, respectively, 81% and 94% in TAn (n=7), and by 49% and 43% in GMn (n=7). Electrical cervical nerve stimulation consistently reduced the isolated tonic discharge in TAn by 66% (n=4, P<0.05) and in GMn by 23% (n=3) when present. Thus the tonic component was more sensitive to inhibition than the rhythmic component of hindlimb muscle nerve activity.

Effect of exercise on lung-perfusion scanning in patients with bronchogenic carcinoma.

The aim of this study was to determine whether perfusion-scintillation scanning, used as a predictive pre-operative index of lung functionality in patients with lung cancer, is affected by the level of pulmonary blood flow (PBF). Twenty patients with primary lung cancer underwent spirometry and a radionuclide-perfusion scan (macroaggregated albumin particles labelled with 99mTechnetium) both at rest and during the last minute of a ramp-like increase in work rate until exhaustion. On average, the perfusion of the lung with the tumour was significantly reduced by the same magnitude at rest and during exercise (mean+/-SD: -9+/-6% versus -10+/-4% of the cardiac output), regardless of the extent of the tumour. However, subject-by-subject analysis revealed that in two patients, a larger decrease in the perfusion of the lung with the tumour was observed during exercise than at rest (-11% and -17%, respectively). This leads to an underestimation of predictive postoperative functional parameters if resting values are used in these patients. The use of perfusion scintigraphy at rest therefore gives a clear picture of the functionality of the lung before resection in most patients requiring surgery.

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.

Neurophysiological basis for neurogenic-mediated articular cartilage anabolism alteration.

This study was designed to investigate the pathways involved in neurogenic-mediated articular cartilage damage triggered by a nonsystemic distant subcutaneous or intra-articular inflammation. The cartilage damage was assessed 24 h after subcutaneous or intra-articular complete Freund's adjuvant (CFA) injection measuring patellar proteoglycan (PG) synthesis (ex vivo [Na(2)(35)SO(4)] incorporation) in 96 Wistar rats. Unilateral subcutaneous or intra-articular injection of CFA induced significant decrease (25-29%) in PG synthesis in both patellae. Chronic administration of capsaicin (50 mg. kg(-1). day(-1) during 4 days), which blunted the normal response of C fiber stimulation, prevented the bilateral significant decrease in cartilage synthesis. Similarly, intrathecal injection of MK-801 (10 nmol/day during 5 days), which blocked the glutamatergic synaptic transmission at the dorsal horn of signal originating in primary afferent C fibers, eliminated the CFA-induced PG synthesis decrease in both patellae. Chemical sympathectomy, induced by guanethidine (12.5 mg. kg(-1). day(-1) during 6 wk), also prevented PG synthesis alteration. Finally, compression of the spinal cord at the T3-T5 level had a similar protective effect on the reduction of [Na(2)(35)SO(4)] incorporation. It is concluded that the signal that triggers articular cartilage synthesis damage induced by a distant local inflammation 1) is transmitted through the afferent C fibers, 2) makes glutamatergic synaptic connections with the preganglionic neurons of the sympathetic system, and 3) involves spinal and supraspinal pathways.

Corticospinal pathway and exercise hyperpnea: lessons from a patient with Arnold Chiari malformation.

The study of a patient with an Arnold Chiari malformation gave us the opportunity to test the hypothesis that the motor cortex contributes significantly to respiratory control during muscular exercise through the corticospinal pathway. The patient was a 25 years old woman who exhibited a severe impairment of the 'automatic' ventilatory control due to a type I Arnold Chiari malformation. Since she never complained of being breathlessness even on exertion, the breath-by-breath ventilatory (VE) and pulmonary gas exchange responses to a three minute bout of constant work rate exercise at 60 W, 90 W and 120 W were studied before then 16 and 23 months after posterior fossa decompression. The VE response to the three different levels of exercise was dramatically blunted so that the expected vertical relationship between PET(CO(2)) and VE during moderate exercise was replaced by an almost horizontal relationship with a slope ranging from 0.15 to 0.17 l/min/Torr. The reduced VE response was associated with a total lack of respiratory sensation during and following the exercise bouts. This abnormal ventilatory response to exercise persisted despite posterior fossa decompression. There was however no evidence of an alteration of the corticospinal pathway. Indeed, not only was there no sign of motor deficit but the patient was able both to mobilize 96% of her expected vital capacity and to voluntarily increase her ventilation to the level expected in a normal subject during exercise. This observation suggests that during exercise, motor control of respiratory muscles via a direct corticospinal pathway does not play a major role in adjusting phrenic motoneuron activity to the magnitude of the motor inputs to the exercising skeletal muscles.

Stimulation of ventilation by normobaric hyperoxia in exercising dogs.

In order to describe the factors which, during hyperoxic exercise, can counteract the chemoreceptor-mediated inhibition of ventilation by O2, minute ventilation (VE) and the pulmonary gas exchange were studied breath-by-breath in four dogs running on a treadmill (5 km x h(-1)) for 10 min during and following exposure to O2 of different durations. We found that a brief inhalation of O2 applied during the steady state of the VE response provoked a reduction in VE by 6.5 +/- 0.9 l x min(-1) whereas hyperoxia applied 2 min before the onset of exercise and maintained for 2.5 min during the running tests had a significantly weaker effect on VE (-1.8 +/- 0.2 l x min(-1), P < 0.05). The rise in pulmonary CO2 output (VCO2) during the prolonged O2 exposure was less than in normoxic exercise leading to a deficit of CO2 eliminated by the lungs of 181 ml. The return to air breathing provoked a rise in VE, which reached within 73 s a much higher level than the control tests (22.9 +/- 3.6 vs. 19.5 +/- 2.2 l x min(-1), P < 0.05); VE then subsided to control levels with a long exponential decline. The CO2 deficit during O2 breathing, was fully compensated after recovery in air within 6 min. No stimulatory effect on ventilation was observed at rest at the cessation of a similar exposure to O2 despite a higher end-tidal PCO2 (+4 +/- 1 mmHg) than in exercise. In conclusion, the stimulatory effect of O2 during exercise can be clearly revealed after recovery in air and seems to operate through a more complex mechanism than that thought to be involved at rest. We propose that the changes in CO2 stores in the exercising muscles could contribute to O2-induced stimulation during exercise, possibly through stimulation of muscle afferents responding to local circulatory changes. Finally, the observation that during continuous dopamine (DA) infusion (5 microg x kg(-1) x min(-1)) the VE response to recovery in air was only a slow decrease, suggests that the arterial chemoreceptors potentiate O2-induced hyperventilation, or that the vascular actions of DA counteract part of the effects provoked by CO2 accumulation in the exercising muscles.

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.

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.

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.

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.

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.

Laryngeal reflex apnea is blunted during and after hindlimb muscle contraction in sheep.

This study was carried out on seven chloralose-anesthetized sheep and was designed to investigate the role of muscular afferent fiber stimulation on the duration of reflex apnea triggered by laryngeal stimulation (LS). In six animals, injection of distilled water onto the laryngeal mucosa provoked a 15.7 +/- 1.0 s (mean +/- SE) apnea associated with a rise in systemic blood pressure (+7 +/- 0.8 Torr). Electrically induced contractions (EIC) of the hindlimb muscles doubled the metabolic rate and ventilation and reduced the duration of the apnea produced by LS to 7.4 +/- 1.0 s (P < 0.01). Apnea duration was still reduced during the first minute after the cessation of EIC (7.2 +/- 1.1 s, P < 0.01) but returned to control after a 5-min recovery period (16.7 +/- 1.6 s). The apnea triggered by LS was also reduced during EIC when the venous return was impeded by occluding the inferior vena cava (5.2 +/- 1.1 s, P < 0.01), despite a profound hypocapnia (20.7 +/- 0.3 Torr). The duration of apnea was not significantly affected (14.2 +/- 1.4 s) by breathing a 6% CO2-14% O2 in N2 gas mixture that roughly mimicked the alveolar gas composition when the apnea turned off. These results suggest that chemical drive has a negligible role in the fast reinitiation of breathing after LS during muscular stimulation. Stimulation of muscle afferent fibers does, however, appear to be a potent source of ventilatory reflexes capable of counteracting the inhibition of breathing resulting from laryngeal stimulation. Conversely, it is postulated that any reduction in somatic afferent traffic during this type of reflex apnea, including that resulting from the LS-induced systemic vasoconstriction, may delay the termination of apnea.