Diaphragm Electromyography Research Articles

Functional Condition Of Inspiratory Muscles In Obstructive Sleep Apnea Patients

Background — Recurrent upper airway obstruction during the sleep is a key feature of obstructive sleep apnea (OSA). In patients with OSA, an intermittent upper airway obstruction yields an increase in inspiratory effort and, consequently, an augmented impact on inspiratory muscles (IM). However, the issue of IM fatigue development in OSA patients is still controversial. Objective — to investigate the functional condition of IM in patients with OSA of various degrees of severity. Material and Methods — Were examined 82 men (30-65 years old), who were distributed among four groups depending on the OSA severity. Cardiorespiratory monitoring was performed to detect OSA. IM electrical activity was detected via surface electromyography (EMG) while holding the breath (during the inhalation). Results — In the course of our study, we detected a reduction in the frequency of EMG of accessory IM, specifically sternocleidomastoid muscle (SCMM) and external intercostal muscles (EIM), accompanied by a simultaneous increase in the amplitude of EMG of SCMM, EIM, and diaphragm (D), while holding the breath (during the inhalation), in the group of patients without OSA. In patients with mild OSA, when holding the breath during the inhalation, the frequency of EMG of SCMM and EIM decreased, while the amplitude of SCMM and EIM increased. In patients with moderate OSA, when holding the breath during the inhalation, a decrease in the frequency of SCMM EMG with a simultaneous increase in the amplitude of the EIM EMG were revealed. Same maneuver in the group of patients with severe OSA yielded no significant changes in the frequency and amplitude of EMG of respiratory muscles. Conclusion — Periodic obstruction of the upper airways with OSA leads to the development of fatigue resistance in accessory IM.

Epidural electrical spinal cord stimulation of the thoracic segments (T2-T5) facilitates respiratory function in patients with complete spinal cord injury

IntroductionPatients with high cervical Spinal Cord Injury (SCI) usually require mechanical ventilation support. Phrenic Nerve Stimulation (PNS) both direct and indirect is the main alternative for these patients to wean off ventilator although PNS has several limitations and phrenic nerve could be also damaged after cervical spinal cord injury. ObjectiveIn this study, we assessed if the spinal cord Epidural Electrical Stimulation (EES) at the segments T2-T5, related to intercostal muscles, can facilitate respiratory function and particularly inspired tidal volume during mechanic ventilation. MethodsTwo patients with a high cervical injury were selected for this study with ethical committee permission and under review board supervision. A phrenic nerve conduction study with diaphragm electromyography (DEMG) was performed before and after trial of EES. ResultsResults demonstrate that EES at T2-T5 substantially increase the inspired volume. The results of this study also demonstrate that EES at spinal segments T2-T5 can bring patients dependent from mechanical ventilation to pressure support (on CPAP), preventing Baro-trauma and other complications related to mechanical ventilation. ConclusionThese findings suggest that tested approach applied alone or in combination with the phrenic nerve stimulation could help to reduce time on mechanical ventilation and related complications.

Neural ventilatory drive decline as a predominant mechanism of obstructive sleep apnoea events

BackgroundIn the classic model of obstructive sleep apnoea (OSA), respiratory events occur with sleep-related dilator muscle hypotonia, precipitating increased neural ventilatory ‘drive’. By contrast, a drive-dependent model has been proposed,...

Compensatory responses to increased mechanical abnormalities in COPD during sleep.

To assess whether night-time increases in mechanical loading negatively impact respiratory muscle function in COPD and whether compensatory increases in inspiratory neural drive (IND) are adequate to stabilize ventilatory output and arterial oxygen saturation, especially during sleep when wakefulness drive is withdrawn. 21 patients with moderate-to-severe COPD and 20 age-/sex-matched healthy controls (CTRL) participated in a prospective, cross-sectional, one-night study to assess the impact of COPD on serial awake, supine inspiratory capacity (IC) measurements and continuous dynamic respiratory muscle function (esophageal manometry) and IND (diaphragm electromyography, EMGdi) in supine sleep. Supine inspiratory effort and EMGdi were consistently twice as high in COPD versus CTRL (p < 0.05). Despite overnight increases in awake total airways resistance and dynamic lung hyperinflation in COPD (p < 0.05; not in CTRL), elevated awake EMGdi and respiratory effort were unaltered in COPD overnight. At sleep onset (non-rapid eye movement sleep, N2), EMGdi was decreased versus wakefulness in COPD (-43 ± 36%; p < 0.05) while unaffected in CTRL (p = 0.11); however, respiratory effort and arterial oxygen saturation (SpO2) were unchanged. Similarly, in rapid eye movement (stage R), sleep EMGdi was decreased (-38 ± 32%, p < 0.05) versus wakefulness in COPD, with preserved respiratory effort and minor (2%) reduction in SpO2. Despite progressive mechanical loading overnight and marked decreases in wakefulness drive, inspiratory effort and SpO2 were well maintained during sleep in COPD. Preserved high inspiratory effort during sleep, despite reduced EMGdi, suggests continued (or increased) efferent activation of extra-diaphragmatic muscles, even in stage R sleep. The COPD data reported herein were secondary data (Placebo arm only) obtained through the following Clinical Trial: "Effect of Aclidinium/Formoterol on Nighttime Lung Function and Morning Symptoms in Chronic Obstructive Pulmonary Disease" ( https://clinicaltrials.gov/ct2/show/NCT02429765 ; NCT02429765).

Evaluation of Diaphragmatic electromyography of cervical spinal cord hemi section in Rat model

Evaluation of Diaphragmatic electromyography of cervical spinal cord hemi section in Rat model

Chapter 2 - Electrophysiological assessment of respiratory function

While the traditional lung function tests are used to assess lung capacity and pulmonary function, they cannot evaluate respiratory driving function and the integrity of the conduction pathway from the central nervous system to the respiratory motor neuron in the spinal cord and to the diaphragm. The inspiratory trigger is sent from the central nervous system through the phrenic nerve and drives the diaphragm to generate inspiratory movement. Therefore, phrenic nerve stimulation and diaphragmatic electromyography are two fundamental methods to assess respiratory function. There are several useful tools to assess respiratory motor system including electrical or magnetic phrenic nerve stimulation, diaphragmatic needle electromyography, and diaphragmatic ultrasound. By these means, physicians can assess current respiratory status in different neurological diseases that affect respiratory muscles, follow-up of the severity of respiratory impairment, help to predict the chance of successfully weaning from ventilatory support, and confirm clinical diagnoses such as diaphragmatic myoclonus. Although some of these tests require special training, applying these neurophysiological assessments in clinical practice is highly recommended.

Evaluation of Correlation Between Surface Diaphragm Electromyography and Airflow Using Fixed Sample Entropy in Healthy Subjects.

In clinic, the acquisition of airflow with nasal prongs, masks, thermistor to monitor respiratory function is more uncomfortable and inconvenience than surface diaphragm electromyography (EMGdi) using electrode pads. The EMGdi with strong electrocardiograph (ECG) interference affect the extraction of its characteristic information. In this work, surface EMGdi and airflow signals of 20 subjects were collected under 5 incremental inspiratory threshold loading protocols from quiet breathing to maximum forced breathing. First, we filtered out the ECG interference in EMGdi based on the combination of stationary wavelet transform and the positioning of ECG to obtain pure EMGdi (EMGdip). Second, the Spearman's rank correlation coefficients between EMGdi and EMGdip quantified by time series fixed sample entropy (fSampEn), root mean square (RMS), and envelope were compared to verify the robustness of the fSampEn to ECG. A comparative analysis of correlation between fSampEn of EMGdi and inspiratory airflow and the correlation between envelope of EMGdip (EMGdie) and inspiratory airflow found that there was no significant difference between the two, indicating the feasibility of using fSampEn to predict airflow. Moreover, fSampEn of EMGdi was used as characteristic parameter to build a quantitative relationship with the airflow by polynomial regression analysis. Mean coefficient of determination of all subjects in any breathing state is greater than 0.88. Finally, nonlinear programming method was used to solve a universal fitting coefficient between fSampEn of EMGdi and airflow for each subject to further evaluate the possibility of using surface EMGdi to monitor and control respiratory activity.

Mechanisms of respiratory depression induced by the combination of buprenorphine and diazepam in rats

BackgroundThe safety profile of buprenorphine has encouraged its widespread use. However, fatalities have been attributed to benzodiazepine/buprenorphine combinations, by poorly understood mechanisms of toxicity. Mechanistic hypotheses include (i) benzodiazepine-mediated increase in brain buprenorphine (pharmacokinetic hypothesis); (ii) benzodiazepine-mediated potentiation of buprenorphine interaction with opioid receptors (receptor hypothesis); and (iii) combined effects of buprenorphine and benzodiazepine on respiratory parameters (pharmacodynamic hypothesis). MethodsWe studied the neuro-respiratory effects of buprenorphine (30 mg kg−1, i.p.), diazepam (20 mg kg−1, s.c.), and diazepam/buprenorphine combination in rats using arterial blood gas analysis, plethysmography, and diaphragm electromyography. Pretreatments with various opioid and gamma-aminobutyric acid receptor antagonists were tested. Diazepam impact on brain 11C-buprenorphine kinetics and binding to opioid receptors was studied using positron emission tomography imaging. ResultsIn contrast to diazepam and buprenorphine alone, diazepam/buprenorphine induced early-onset sedation (P<0.05) and respiratory depression (P<0.001). Diazepam did not alter 11C-buprenorphine brain kinetics or binding to opioid receptors. Diazepam/buprenorphine-induced effects on inspiratory time were additive, driven by buprenorphine (P<0.0001) and were blocked by naloxonazine (P<0.01). Diazepam/buprenorphine-induced effects on expiratory time were non-additive (P<0.001), different from buprenorphine-induced effects (P<0.05) and were blocked by flumazenil (P<0.01). Diazepam/buprenorphine-induced effects on tidal volume were non-additive (P<0.01), different from diazepam- (P<0.05) and buprenorphine-induced effects (P<0.0001) and were blocked by naloxonazine (P<0.05) and flumazenil (P<0.05). Compared with buprenorphine, diazepam/buprenorphine decreased diaphragm contraction amplitude (P<0.01). ConclusionsPharmacodynamic parameters and antagonist pretreatments indicate that diazepam/buprenorphine-induced respiratory depression results from a pharmacodynamic interaction between both drugs on ventilatory parameters.

Ampakines Stimulate Diaphragm Activity after Spinal Cord Injury.

Respiratory compromise after cervical spinal cord injury (SCI) is a leading cause of mortality and morbidity. Most SCIs are incomplete, and spinal respiratory motoneurons as well as proprio- and bulbospinal synaptic pathways provide a neurological substrate to enhance respiratory output. Ampakines are allosteric modulators of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors, which are prevalent on respiratory neurons. We hypothesized that low dose ampakine treatment could safely and effectively increase diaphragm electromyography (EMG) activity that has been impaired as a result of acute- or sub-acute cervical SCI. Diaphragm EMG was recorded using chronic indwelling electrodes in unanesthetized, freely moving rats. A spinal hemi-lesion was induced at C2 (C2Hx), and rats were studied at 4 and 14 days post-injury during room air breathing and acute respiratory challenge accomplished by inspiring a 10% O2, 7% CO2 gas mixture. Once a stable baseline recording was established, one of two different ampakines (CX717 or CX1739, 5 mg/kg, intravenous) or a vehicle (2-hydroxypropyl-beta-cyclodextrin [HPCD]) was delivered. At 4 days post-injury, both ampakines increased diaphragm EMG output ipsilateral to C2Hx during both baseline breathing and acute respiratory challenge. Only CX1739 treatment also led to a sustained (15 min) increase in ipsilateral EMG output. At 14 days post-injury, both ampakines produced sustained increases in ipsilateral diaphragm EMG output and enabled increased output during the respiratory challenge. We conclude that low dose ampakine treatment can increase diaphragm EMG activity after cervical SCI, and therefore may provide a pharmacological strategy that could be useful in the context of respiratory rehabilitation.

Effects of the Elevation Training Mask® 2.0 on dyspnea and respiratory muscle mechanics, electromyography, and fatigue during exhaustive cycling in healthy humans

ObjectivesExamine the effects of the Elevation Training Mask® 2.0 (ETM) on dyspnea, and respiratory muscle function and fatigue during exercise. DesignRandomized crossover. Methods10 healthy participants completed 2 time-to-exhaustion (TTE) cycling tests while wearing the ETM or under a sham control condition. During the sham, participants were told they were breathing air equivalent to “9000 ft” (matched to the selected resistance valves on the ETM according to the manufacturer), but they were breathing room air. Dyspnea and leg discomfort were assessed using the modified 0–10 category-ratio Borg scale. Qualitative dyspnea descriptors at peak exercise were selected from a list of 15. Crural diaphragmatic electromyography (EMGdi) and transdiaphragmatic pressure (Pdi) were measured via a multipair esophageal electrode balloon catheter. Participants performed maximal respiratory maneuvers before and after exercise to estimate the degree of respiratory muscle fatigue. ResultsExercise with the ETM resulted in a significant decrease in TTE (p = 0.015), as well as increased dyspnea at baseline (p = 0.032) and during the highest equivalent submaximal exercise time (p = 0.0001). The increase in dyspnea with the ETM was significantly correlated with the decrease in exercise time (r = 0.73, p = 0.020). EMGdi and Pdi were significantly increased with the ETM at all time points (all p < 0.05). There was a significant increase in the selection frequency of “my breath does not go in all the way” at peak exercise with the ETM (p = 0.02). The ETM did not induce respiratory muscle fatigue. ConclusionsExercising with the ETM appears to decrease exercise performance, in part, by increasing the sensation of dyspnea.

The control of respiratory pressures and neuromuscular activation to increase force production in trained martial arts practitioners.

The mechanisms that explain the ability of trained martial arts practitioners to produce and resist greater forces than untrained individuals to aid combat performance are not fully understood. We investigated whether the greater ability of trained martial arts practitioners to produce and resist forces was associated with an enhanced control of respiratory pressures and neuromuscular activation of the respiratory, abdominal, and pelvic floor musculature. Nine trained martial arts practitioners and nine untrained controls were instrumented with skin-surface electromyography (EMG) on the sternocleidomastoid, rectus abdominis, and the group formed by the transverse abdominal and internal oblique muscles (EMGtra/io). A multipair oesophageal EMG electrode catheter measured gastric (Pg), transdiaphragmatic (Pdi), and oesophageal (Pe) pressures and EMG of the crural diaphragm (EMGdi). Participants performed Standing Isometric Unilateral Chest Press (1) and Standing Posture Control (2) tasks. The trained group produced higher forces normalised to body mass2/3 (0.033 ± 0.01 vs. 0.025 ± 0.007N/kg2/3 mean force in Task 1), lower Pe, and higher Pdiin both tasks. Additionally, they produced higher Pg (73 ± 42 vs. 49 ± 19 cmH2O mean Pg) and EMGtra/ioin Task 1 and higher EMGdiin Task 2. The onset of Pgwith respect to the onset of force production was earlier, and the relative contributions of Pg/Pe and Pdi/Pe were higher in the trained group in both tasks. Our findings demonstrate that trained martial arts practitioners utilised a greater contribution of abdominal and diaphragm musculature to chest wall recruitment and higher Pdi to produce and resist higher forces.

Pulmonary Effects of Sustained Periods of High-G Acceleration Relevant to Suborbital Spaceflight.

AbstractBACKGROUND: Members of the public will soon be taking commercial suborbital spaceflights with significant Gx (chest-to-back) acceleration potentially reaching up to 6 Gx. Pulmonary physiology is gravity-dependent and is likely to be affected, which may have clinical implications for medically susceptible individuals.METHODS: During 2-min centrifuge exposures ranging up to 6 Gx, 11 healthy subjects were studied using advanced respiratory techniques. These sustained exposures were intended to allow characterization of the underlying pulmonary response and did not replicate actual suborbital G profiles. Regional distribution of ventilation in the lungs was determined using electrical impedance tomography. Neural respiratory drive (from diaphragm electromyography) and work of breathing (from transdiaphragmatic pressures) were obtained via nasoesophageal catheters. Arterial blood gases were measured in a subset of subjects. Measurements were conducted while breathing air and breathing 15 oxygen to simulate anticipated cabin pressurization conditions.RESULTS: Acceleration caused hypoxemia that worsened with increasing magnitude and duration of Gx. Minimum arterial oxygen saturation at 6 Gx was 86 1 breathing air and 79 1 breathing 15 oxygen. With increasing Gx the alveolar-arterial (A-a) oxygen gradient widened progressively and the relative distribution of ventilation reversed from posterior to anterior lung regions with substantial gas-trapping anteriorly. Severe breathlessness accompanied large progressive increases in work of breathing and neural respiratory drive.DISCUSSION: Sustained high-G acceleration at magnitudes relevant to suborbital flight profoundly affects respiratory physiology. These effects may become clinically important in the most medically susceptible passengers, in whom the potential role of centrifuge-based preflight evaluation requires further investigation.Pollock RD, Jolley CJ, Abid N, Couper JH, Estrada-Petrocelli L, Hodkinson PD, Leonhardt S, Mago-Elliott S, Menden T, Rafferty G, Richmond G, Robbins PA, Ritchie GAD, Segal MJ, Stevenson AT, Tank HD, Smith TG. Pulmonary effects of sustained periods of high-G acceleration relevant to suborbital spaceflight. Aerosp Med Hum Perform. 2021; 92(7):633641.

Sleep increases leaks and asynchronies during home noninvasive ventilation: a polysomnographic study.

In patients treated with noninvasive ventilation, sleep-related breathing changes can modify patient-ventilator interactions, which could reduce its effectiveness. The aim of this prospective observational study was to determine the impact of sleep/wake state on leaks, upper airway obstructive events, and asynchronies in patients treated by long-term noninvasive ventilation. Stable patients adapted to noninvasive ventilation were considered for nocturnal polysomnography. Unintentional leaks, upper airway obstructive events, and asynchronies were compared between sleep and awake periods. Twenty-eight patients were enrolled. Underlying diagnoses were neuromuscular disease (n = 11), chest wall disease (n = 8), and obesity-hypoventilation (n = 9). Leaks were more frequent in sleep than in awake periods, with a median of 10% (interquartile range [IQR], 0%-75%) vs 1% (IQR, 0%-9%) of time (P < .001), respectively. During sleep, asynchronies with and without associated leak affected 27% of breaths (IQR, 16%-39%) compared with non-leak-related asynchronies that were recorded in 8% (IQR, 3%-25%) of breaths (P < .001). Asynchronies affecting more than 10% of total breaths were more frequent in sleep (25 patients, 89%) than in awake time (8 patients, 29%; P = .25). Eleven patients (39%) presented with 5 or more upper airway obstructive events without reduction in ventilatory drive per hour of sleep. In patients adapted to home noninvasive ventilation, leaks, asynchronies, and upper airway obstructive events are frequent during the night and are concentrated in sleep periods. Asynchronies are often associated with leaks. These findings may have clinical implications considering that in patients with low sleep efficiency respiratory events could be underestimated if sleep is not evaluated. Martí S, Ferré A, Sampol G, etal. Sleep increases leaks and asynchronies during home noninvasive ventilation: a polysomnographic study. J Clin Sleep Med. 2022;18(1):225-233.

Intermittent Hypoxia Induces Greater Functional Breathing Motor Recovery as a Fixed Rather Than Varied Duration Treatment after Cervical Spinal Cord Injury in Rats.

Intermittent hypoxia treatment (IH) has been shown to improve respiratory function in both pre-clinical animal models and human subjects following spinal cord injury (SCI), historically consisting of alternating and equal intervals of hypoxic and normoxic exposure. We describe such a procedure as fixed duration IH (FD-IH) and modulation of its severity, intermittency, and post-injury time-point of application differentially affects expression of breathing motor plasticity. As such, the established IH protocol exhibits similarity to instrumental conditioning and can be described as behavioral training through reinforcement. Findings from the field of operant conditioning, a form of more advanced learning, inspire the consideration that FD-IH protocols may be improved through exchanging fixed for varied durations of hypoxia between reinforcement. Thus, we hypothesized that varied duration intermittent hypoxia treatment (VD-IH) would induce greater breathing motor recovery ipsilateral to injury than FD-IH after cervical SCI in rats. To test this hypothesis, we treated animals with VD-IH or FD-IH for 5 days at 1 week and at 8 weeks following cervical SCI, then assessed breathing motor output by diaphragm electromyography (EMG) recording, and compared between groups. At 1 week post-injury, VD-IH-exposed animals trended slightly toward exhibiting greater levels of respiratory recovery in the hemidiaphragm ipsilateral to lesion than did FD-IH-treated animals, but at 8 weeks FD-IH produced significantly greater respiratory motor output than did VD-IH. Thus, these results identify a novel sensitivity of respiratory motor function to variations in the IH protocol that may lead to development of more effective treatments following SCI.

Comparison of the effects between low-level assisted ventilation and T-piece method on respiratory mechanics during weaning of mechanically ventilated patients

To compare the difference of low-level assisted ventilation and T-piece method on respiratory mechanics of patients with invasive mechanical ventilation during spontaneous breathing trial (SBT) within 3 days before extubation. A retrospective observational study was conducted. Twenty-five patients with difficulty in weaning or delayed weaning from invasive mechanical ventilation who were admitted to department of critical care medicine of the First Affiliated Hospital of Guangzhou Medical University from December 2018 to June 2020, and were in stable condition and entered the weaning stage after more than 72 hours of invasive mechanical ventilation were studied. A total of 119 cases of respiratory mechanical indexes were collected, which were divided into the low-level assisted ventilation group and the T-piece group according to the ventilator method and parameters used during the data collection. The different ventilation modes related respiratory mechanics indexes such as the esophageal pressure (Pes), the gastric pressure (Pga), the transdiaphragmatic pressure (Pdi), the maximum Pdi (Pdimax), Pdi/Pdimax ratio, the esophageal pressure-time product (PTPes), the gastric pressure-time product (PTPga), the transdiaphragmatic pressure-time product (PTPdi), the diaphragmatic electromyography (EMGdi), the maximum diaphragmatic electromyography (EMGdimax), PTPdi/PTPes ratio, Pes/Pdi ratio, the inspiratory time (Ti), the expiratory time (Te) and the total time respiratory cycle (Ttot) at the end of monitoring were recorded and compared between the two groups. Compared with the T-piece group, Pes, PTPes, PTPdi/PTPes ratio, Pes/Pdi ratio and Te were higher in low-level assisted ventilation group [Pes (cmH2O, 1 cmH2O = 0.098 kPa): 2.84 (-1.80, 5.83) vs. -0.94 (-8.50, 2.06), PTPes (cmH2O×s×min-1): 1.87 (-2.50, 5.93) vs. -0.95 (-9.71, 2.56), PTPdi/PTPes ratio: 0.07 (-1.74, 1.65) vs. -1.82 (-4.15, -1.25), Pes/Pdi ratio: 0.17 (-0.43, 0.64) vs. -0.47 (-0.65, -0.11), Te (s): 1.65 (1.36, 2.18) vs. 1.33 (1.05, 1.75), all P < 0.05], there were no significant differences in Pga, Pdi, Pdimax, Pdi/Pdimax ratio, PTPga, PTPdi, EMGdi, EMGdimax, Ti and Ttot between the T-piece group and the low-level assisted pressure ventilation group [Pga (cmH2O): 6.96 (3.54,7.60) vs. 7.74 (4.37, 11.30), Pdi (cmH2O): 9.24 (4.58, 17.31) vs. 6.18 (2.98, 11.96), Pdimax (cmH2O): 47.20 (20.60, 52.30) vs. 29.95 (21.50, 47.20), Pdi/Pdimax ratio: 0.25 (0.01, 0.34) vs. 0.25 (0.12, 0.41), PTPga (cmH2O×s×min-1): 7.20 (2.54, 9.97) vs. 7.97 (5.74, 13.07), PTPdi (cmH2O×s×min-1): 12.15 (2.95, 19.86) vs. 6.87 (2.50, 12.63), EMGdi (μV): 0.05 (0.03, 0.07) vs. 0.04 (0.02, 0.06), EMGdimax (μV): 0.07 (0.05, 0.09) vs. 0.07 (0.04, 0.09), Ti (s): 1.20 (0.95, 1.33) vs. 1.07 (0.95, 1.33), Ttot (s): 2.59 (2.22, 3.09) vs. 2.77 (2.35, 3.24), all P > 0.05]. When mechanically ventilated patients undergo SBT, the use of T-piece method increases the work of breathing compared with low-level assisted ventilation method. Therefore, long-term use of T-piece should be avoided during SBT.

Regulation of Coronary Blood Flow by the Carotid Body Chemoreceptors in Ovine Heart Failure.

Carotid bodies (CBs) are peripheral chemoreceptors, which are primary sensors of systemic hypoxia and their activation produces respiratory, autonomic, and cardiovascular adjustments critical for body homeostasis. We have previously shown that carotid chemoreceptor stimulation increases directly recorded cardiac sympathetic nerve activity (cardiac SNA) which increases coronary blood flow (CoBF) in conscious normal sheep. Previous studies have shown that chemoreflex sensitivity is augmented in heart failure (HF). We hypothesized that carotid chemoreceptor stimulation would increase CoBF to a greater extent in HF than control sheep. Experiments were conducted in conscious HF sheep and control sheep (n = 6/group) implanted with electrodes to record diaphragmatic electromyography (dEMG), flow probes to record CoBF as well as arterial pressure. There was a significant increase in mean arterial pressure (MAP), CoBF and coronary vascular conductance (CVC) in response to potassium cyanide (KCN) in both groups of sheep. To eliminate the effects of metabolic vasodilation, the KCN was repeated while the heart was paced at a constant level. In this paradigm, the increase in CoBF and CVC was augmented in the HF group compared to the control group. Pre-treatment with propranolol did not alter the CoBF or the CVC increase in the HF group indicating this was not mediated by an increase in cardiac sympathetic drive. The pressor response to CB activation was abolished by pre-treatment with intravenous atropine in both groups, but there was no change in the CoBF and vascular conductance responses. Our data suggest that in an ovine model of HF, carotid body (CB) mediated increases in CoBF and CVC are augmented compared to control animals. This increase in CoBF is mediated by an increase in cardiac SNA in the control group but not the HF group.

Mechanisms of Exertional Dyspnea in Patients with Mild COPD and a Low Resting DLCO

Patients with mild chronic obstructive pulmonary disease (COPD) and lower resting diffusing capacity for carbon monoxide (DLCO) often report troublesome dyspnea during exercise although the mechanisms are not clear. We postulated that in such individuals, exertional dyspnea is linked to relatively high inspiratory neural drive (IND) due, in part, to the effects of reduced ventilatory efficiency. This cross-sectional study included 28 patients with GOLD I COPD stratified into two groups with (n = 15) and without (n = 13) DLCO less than the lower limit of normal (<LLN; Global Lung Function Initiative criteria) and 16 healthy controls. We compared dyspnea (Borg scale), IND (by diaphragm electromyography), ventilatory equivalent for CO2 (V̇ E/V̇CO2), and respiratory mechanics during incremental cycle exercise in the three groups. Spirometry and resting lung volumes were similar between COPD groups. During exercise, dyspnea, IND and V̇ E/V̇CO2 were higher at equivalent work rates (WR) in the DLCO<LLN group compared with the other two groups (all p < 0.05). In patients with DLCO<LLN, severe respiratory mechanical constraints, indicated by end-inspiratory lung volume of approximately 90% of total lung capacity, occurred at a lower WR than the other two groups (p < 0.05). The dyspnea/IND relationship was similar across groups; therefore, the increased dyspnea at a standardized WR in the low DLCO<LLN group reflected the higher corresponding IND. Higher dyspnea ratings in patients with mild COPD and DLCO<LLN were associated with higher IND and V̇ E/V̇CO2 at a given work rate. Higher ventilatory requirements in the DLCO<LLN group accelerated dynamic mechanical abnormalities earlier in exercise, further increasing IND and dyspnea.

Optogenetic diaphragm muscle activation in mice

Breathing impairments are the leading cause of morbidity and mortality across all forms of neuromuscular disease. When voluntarily diaphragm activation is insufficient, clinical options include mechanical ventilation or electrical stimulation via intramuscular wires. Here we tested the hypothesis that an optogenetic approach would enable light to serve as an adequate stimulus for inducing diaphragm contraction to sustain breathing during neuromuscular impairments. Wild type, C57/bl6 mice (n=10) received an intrapleural injection of an adeno-associated virus (AAV) construct containing the transgene for the light activated ion channel channelrhodopsin-2 (34 µl, AAV9-ACAGW-ChR2-mVenus, titer: 1.8x1013 vg/ml). After allowing 8-10 weeks for transgene expression, electromyography (EMG) and pneumotachography were measured during spontaneous breathing under isoflurane anesthesia. To test optogenetic activation, a high-power, 472 nm, micro-LED was directed at the inferior diaphragm surface. Stimulus pulse durations (PD) ranging from 0.1-1.0 ms and interpulse intervals (IPI) of 0.1, 0.5, 1.0, and 5.0 ms were tested. Directing light at the diaphragm evoked clearly discernable EMG activity in 10 of 10 animals treated with the AAV9 vector. The 5 ms IPI induced the largest EMG activation, occurring at a level comparable to endogenous diaphragm bursts. Stimulation with a 0.1 ms PD / 0.1 ms IPI, and 1 ms PD / 5 ms IPI both were able to generate robust increases in respiratory airflow. Optogenetic stimulation was repeated following an acute unilateral phrenic nerve section which abolished ipsilateral diaphragm EMG. The contralateral diaphragm inspiratory EMG signal was then used to “trigger” the light stimulation of the paralyzed hemi-diaphragm. This procedure enabled a robust, bilateral diaphragm activation in all mice that received the unilateral phrenicotomy (n= 5). Histology confirmed robust expression of mVenus in diaphragm myofibers in mice treated with the vector. Directing light at the diaphragm of non-AAV treated control mice caused no discernible impact on diaphragm EMG activity (n=2). We conclude that intrapleural AAV9-ACAGW-ChR2-mVenus injection effectively drives transgene expression in the diaphragm, and this approach can be used to enable robust optogenetic activation of this critically important respiratory muscle. To our knowledge, this is the first demonstration of light activation of the diaphragm, and this success opens up new directions for therapeutic targeting of diaphragm paralysis and respiratory insufficiency.

Selective Inhibition of Pre‐Bötzinger Neurons Projecting to the Facial Nucleus Decreases Nasofacial Activity Without Affecting Breathing

The pre-Bötzinger Complex (preBötC) is a major generator of respiratory rhythm and is crucial for respiratory modulation of orofacial behaviours. The nasolabialis profundus (NLP) is a facial muscle located in the rostral part of the rat snout that causes vibrissae protraction. NLP motoneurons receive direct input from the preBötC, but the identity of the responsible preBötC neurons is not fully elucidated. The aim of this study was to identify this preBötC neurons sub-group based on their axonal projections and physiological function. To achieve this, we developed a cre-recombinase (Cre) dependent adeno-associated virus (AAV) that expressed the light-activated chloride channel (GtACR2) predominantly in the soma (AAV-DIO-GtACR2-muGFP-Kv2.1). To validate this new AAV, it was injected with AAV-Cre-TdTomato into the preBötC of Sprague-Dawley rats (60-80 g, n=2). Three weeks later, rats were anesthetised with intravenous urethane, and blood pressure (BP), heart rate (HR), diaphragm (dEMG) and NLP electromyography (EMG) were recorded. Optical fibers were bilaterally implanted into the preBötC, and 470 nm light (50 Hz, 5 ms pulse, 20 mW) delivered for 30 s to photoinhibit neurons. The results are expressed as mean ± SD. We observed effective Cre recombination, resulting in GtACR2-muGFP expression in the preBötC. During photoinhibition of preBötC, dEMG initially ceased, but this did not last for the entire photoinhibition period (8 ± 2 s); HR decreased (ΔHR: -10 ± 1.2% from baseline); and BP decreased during apnea (ΔMAP: -10 ± 10 % from baseline), but returned toward baseline levels as breathing resumed. Under baseline conditions, activity of the NLP occurred almost exclusively during inspiration, with a limited pre-I component activity. All NLP EMG activity ceased during initial photoinhibition and then returned with activity in the dEMG. During the photoinhibition, when breathing had resumed, we observed an enlargement of the duration of the pre-I component of NLP (0.27 ± 0.02 vs. baseline: 0.06 ± 0.0004 s). To selectively inhibit preBötC neurons that project to the NLP motoneurons, we injected a retrogradely transporting AAV expressing Cre (AAVrg-mCherry-Cre) into the lateral edge of the 7n, and AAV-DIO-GtACR2-muGFP-Kv2.1 into the preBötC (n=2). Three weeks later, we performed the same experimental protocol as described above. Postmortem histology showed strong muGFP expression in a sub-group of preBötC neurons. Selective photoinhibition of these preBötC neurons that project to the 7n neurons reduced NLP activity (Δ amplitude: -42 ± 6% from baseline) and the pre-I component (0.03 ± 0.02 vs. baseline: 0.08 ± 0.001 s s), without affecting breathing, BP or HR. In conclusion, beyond validating a new optogenetic tool, our results suggest that separate populations of preBötC neurons, with coordinated respiratory rhythmicity, drive dEMG and NLP EMG activity.

Neuromuscular efficiency is impaired during exercise in COPD patients

Aimto analyze respiratory and peripheral neuromuscular efficiency during exercise in COPD. MethodsCOPD patients (VEF1 = 39.25 ± 13.1 %) were paired with healthy subjects. It was performed cardiopulmonary exercise test with simultaneously electromyography (EMG). Respiratory neuromuscular efficiency was determined by relationship between tidal volume and diaphragm EMG. Peripheral neuromuscular efficiency was determined by relationship between power output and vastus lateralis EMG. ResultsHealthy subjects presented higher respiratory neuromuscular efficiency at moderate, heavy and maximum exercise intensities compared to COPD (p < 0.05). Healthy subjects presented higher peripheral neuromuscular efficiency at light, moderate, heavy and maximum exercise intensities compared to COPD (p < 0.001). Dynamic hyperinflation presented correlation with respiratory and peripheral neuromuscular efficiency (r = −0.73 and r = −0.76, p < 0.001). ConclusionCOPD patients have lower respiratory neuromuscular efficiency at moderate exercise intensity and lower peripheral neuromuscular efficiency at light exercise intensity. Dynamic hyperinflation affects respiratory and peripheral neuromuscular efficiency.