Laryngeal stimulation by an acid solution in the pre-term lamb.

In a mature organism, the contact between various liquids and the laryngeal mucosa triggers lower airway protective responses (cough, swallowing, arousal). These laryngeal chemoreflexes (LCR) are essential for preventing aspiration. In contrast, previous studies showed that LCR are responsible for apnea and bradycardia in the neonatal mammal. Consequently, LCR, especially when triggered by acid gastrolaryngeal reflux, are deemed responsible for some apneas of prematurity and many life-threatening events of infancy and, probably, for some cases of sudden infant death syndrome. Recently, we have revisited LCR in full-term lambs during quiet sleep. Our results showed that the LCR triggered by HCl (pH 2), mimicking the acid component of an acid gastro-oesophageal reflux, were consistently like the mature LCR reported in adult mammals, without significant apneas and bradycardias (St-Hilaire 2005). These results prompted us to question whether premature birth alters LCR. Results show that LCR triggered in pre-term lambs by both saline and HCl are much more marked and clinically relevant than the ones observed in full-term lambs. Indeed, life-threatening responses to HCl, including repetitive apneas for more than 90 seconds, severe desaturation and bradycardia, were observed in 2 lambs at postnatal day 7 (D7). In addition, LCR were significantly blunted at D14. In conclusion, HCl can trigger potentially dangerous LCR in pre-term lambs at D7, suggesting that LCR in response to acid gastrolaryngeal refluxes are likely involved in some apnea/bradycardia/desaturation in pre-term infants, before they reach a post-conceptional age equivalent to full gestation.

CPAP inhibits non-nutritive swallowing through stimulation of bronchopulmonary receptors.

While swallowing and respiratory problems are among the most frequent disorders encountered in neonates, the interrelationships between both functions are not completely understood. This is especially true for non-nutritive swallowing (NNS), which fulfills the important function of clearing upper airways from both local secretions and liquids refluxed from the stomach. Recently, we showed that nasal CPAP inhibits NNS during quiet sleep in the newborn lamb (Samson, St-Hilaire, Nsegbe, Reix, Moreau-Bussière and Praud 2005). The present study was aimed at testing the hypothesis that NNS inhibition is eliminated when CPAP is directly administered through a tracheostomy, thus eliminating reflexes originating from upper airway receptors. Results show that both nasal and tracheal CPAP 6cm H2O similarly inhibit total NNS during quiet sleep, thus suggesting that the inhibiting effect of nasal CPAP on NNS is mainly mediated through bronchopulmonary mechanical receptors with minimal participation of the upper airways.

Effects of hypercapnia on non-nutritive swallowing in newborn lambs.

The purpose of this study was to determine the effect of hypercapnia on non-nutritive swallowing (NNS) frequency and on NNS-breathing coordination in newborn lambs. Six lambs were chronically instrumented for recording electroencephalogram, eye movements, diaphragm and thyroarytenoid muscle activity, nasal airflow and electrocardiogram. Each lamb was placed in a Plexiglas chamber and exposed to a hypercapnic gas mixture (21% O2, 5% CO2). Polysomnographic recordings were conducted in non-sedated lambs using a custom-designed radiotelemetry system. Results show that hypercapnia increased NNS frequency in all three states of alertness (p < 0.0001 to 0.03), through a specific increase in ie-type NNS. Causal mechanisms and potential consequences of such observations on aspirations and apneas, as well as on swallowing maturation, will require further studies.

Effects of hypoxia and hypercapnia on nonnutritive swallowing in newborn lambs.

The aim of the present study was to investigate the effect of hypercapnia and hypoxia on apnea and nonnutritive swallowing (NNS) frequency, as well as on the coordination between NNS and phases of the respiratory cycle in newborn lambs, while taking into account the potential effects of states of alertness. Six lambs were chronically instrumented for recording electroencephalogram, eye movements, diaphragm and thyroarytenoid muscle (a glottal adductor) activity, nasal airflow, and electrocardiogram. Polysomnographic recordings were performed in nonsedated lambs exposed to air (control), 10% O(2), and 5% CO(2) in a random order at 3, 4, and 5 days of age. Although hypercapnia decreased apnea frequency in wakefulness and active sleep (P = 0.002 vs. air and hypoxia), hypoxia had no significant effect on apnea. In addition, although hypercapnia increased NNS frequency during wakefulness and quiet sleep (P < 0.005 vs. air and hypoxia), hypoxia tended to decrease NNS frequency. Finally, only hypercapnia altered NNS-breathing coordination by increasing NNS at the transition from inspiration to expiration (ie-type NNS; P < 0.001 vs. air and hypoxia). In conclusion, whereas hypercapnia increases overall NNS frequency by specifically increasing ie-type NNS, hypoxia has the inverse tendency. Results were identical in all three states of alertness.

Postnatal maturation of laryngeal chemoreflexes in the preterm lamb.

Laryngeal chemoreflexes (LCR) are triggered by the contact of liquids with the laryngeal mucosa. In the mature organism, LCR trigger lower airway protective responses (coughing, effective swallowing, and arousal) to prevent aspiration. General belief holds that LCR are responsible for apnea and bradycardia in the newborn mammal, including humans. Our laboratory has recently shown that LCR in full-term lambs are consistently analogous to the mature LCR reported in adult mammals, without significant apneas and bradycardias (St-Hilaire M, Nsegbe E, Gagnon-Gervais K, Samson N, Moreau-Bussiere F, Fortier PH, and Praud J-P. J Appl Physiol 98: 2197-2203, 2005). The aim of the present study was to assess LCR in nonsedated, newborn preterm lambs born at 132 days of gestation (term = 147 days). The preterm lambs were instrumented for recording glottal adductor electromyogram, electroencephalogram, eye movements, heart rate, respiration, and oximetry. A chronic supraglottal catheter was used for injecting 0.5 ml of saline, distilled water, and HCl (pH 2) during quiet sleep, active sleep, and wakefulness on postnatal days 7 (D7) and 14 (D14). Laryngeal stimulation by water or HCl on D7 induced significant apneas, bradycardia, and desaturation, which, at times, appeared potentially life-threatening. No significant apneas, bradycardias, or desaturation were observed on D14. No consistent effects of sleep state could be shown in the present study. In conclusion, laryngeal stimulation by liquids triggers potentially dangerous LCR in preterm lambs on D7, but not on D14. It is proposed that maturation of the LCR between D7 and D14 is partly involved in the disappearance of apneas/bradycardias of prematurity with postnatal age.

C-fiber blockade influence on non-nutritive swallowing in full-term lambs.

Systemic C-fiber blockade (CFB) has been reported to inhibit induced swallowing in adult guinea pigs. The aim of this study was to investigate the effects of CFB on spontaneous, non-nutritive swallowing (NNS) frequency and NNS-respiration coordination in the neonatal period. Seven CFB lambs and seven control lambs aged 2+/-1 days were chronically instrumented for recording electroencephalogram, eye movements, diaphragm EMG, thyroarytenoid muscle EMG, nasal airflow and electrocardiogram. Polysomnographic recordings were performed in non-sedated lambs, using radiotelemetry transmission. CFB lambs spent more time in active sleep than controls (p=0.02). Frequency of non-nutritive swallowing was not different in CFB and control lambs, whatever the state of alertness. In addition, CFB did not disrupt the overall respiratory-swallowing coordination, inspiratory-related NNS being the most frequent and expiratory-related NNS the least in both CFB and control lambs. Further analyses revealed that CFB had no effect on baseline respiratory and heart rate, and apnea and sigh frequency, whatever the state of alertness. Our results suggest that, in the neonatal period, C-fibers are not involved in NNS frequency and have no influence on the overall respiratory-swallowing coordination.

Monitoring pulse oximetry via radiotelemetry in freely-moving lambs.

This study was aimed at validating the use of a custom-made wireless pulse oximeter in freely moving lambs, using radiotelemetry transmission. First, measurements obtained simultaneously using the new, wireless oximeter and a standard commercially-available pulse oximeter (Nonin 8500) were compared in five lambs during 5min episodes of normoxia, hypoxia and hyperoxia. Correlation between the two oximeters for both SpO(2) and heart rate was very good, regardless of oxygenation conditions. Secondly, the capabilities of our device were assessed during more than 45h of polysomnographic recordings in seven lambs. According to the plethysmographic pulse waveform, reliable SpO(2) values were obtained in more than 85% of recording time. Multiple decreases in SpO(2) were readily observed after spontaneous apneas in preterm lambs. It is concluded that our wireless pulse oximeter performs as reliably as a standard pulse oximeter for monitoring SpO(2) variations in lambs, and offers new perspectives for researchers interested in continuous monitoring of oxygenation throughout sleep stages and wakefulness.

Closure of the uterine incision with one or two layers after caesarean section: a randomized controlled study in sheep.

OBJECTIVE: To compare the quality of the uterine scar with one or two layer closure after caesarean section by studying biomechanical and pathological properties of the scar. METHODS: A randomized controlled trial performed on eight term pregnant ewes assigned into two groups during caesarean according to type of uterine closure: single-layer or double-layer. Hysterectomy was performed 8 months after caesarean delivery. Tensile strength of all scars and of unscarred myometrium was measured. Pathological properties of the scars were analyzed histologically. RESULTS: The force required to reach the yield point was similar between scarred and unscarred myometrium (p=0.96), and between the scars in single-layer and double-layer closure groups (p=0.65). There was a significant increase in fibrosis width on the superficial part of the uterus in the double-layer closure group compared to the single-layer group (p=0.02). CONCLUSIONS: Double-layer uterine closure modified wound healing without significant change in biomechanical properties.

Effects of postnatal environmental tobacco smoke on non-nutritive swallowing-breathing coordination in newborn lambs.

While prenatal environmental tobacco smoke (ETS) exposure is a well-known risk factor for sudden infant death syndrome, the effect of postnatal ETS exposure is less clear. The objective of this study was to investigate the effect of postnatal ETS exposure on non-nutritive swallowing (NNS) and NNS-breathing coordination, which are crucial to prevent aspiration related-cardiorespiratory events. Eighteen newborn lambs (6 per group) were randomly exposed to either 10 cigarettes/day, 20 cigarettes/day or room air for 15 days. Lambs were instrumented for recording states of alertness, swallowing, electrocardiogram and breathing; recordings were performed in non-sedated lambs at the end of ETS exposure. Urinary cotinine/creatinine ratio confirmed relevant real-life exposure. Postnatal ETS exposure had no effect on NNS frequency but tended to decrease inspiratory NNS (p=0.07) during quiet sleep. No effect on respiratory or heart rate (p>0.6), apnea index (p=0.2) or sleep states (p=0.3) was observed. In conclusion, postnatal ETS exposure in lambs had only mild effects on NNS-breathing coordination.

Influence of hypoxia and hypercapnia on sleep state-dependent heart rate variability behavior in newborn lambs.

STUDY OBJECTIVES: Although hypercapnia and/or hypoxia are frequently present during chronic lung disease of infancy and have also been implicated in sudden infant death syndrome (SIDS), their effect on cardiac autonomic regulation remains unclear. The authors' goal is to test that hypercapnia and hypoxia alter sleep-wake cycle-dependent heart rate variability (HRV) in the neonatal period. DESIGN: Experimental study measuring HRV during sleep states in lambs randomly exposed to hypercapnia, hypoxia, or air. SETTING: University center for perinatal research in ovines (Sherbrooke, Canada). INSERM-university research unit for signal processing (Rennes, France). PARTICIPANTS: Six nonsedated, full-term lambs. INTERVENTIONS: Each lamb underwent polysomnographic recordings while in a chamber flowed with either air or 21% O(2) + 5% CO(2) (hypercapnia) or 10% O(2) + 0% CO(2) (hypoxia) on day 3, 4, and 5 of postnatal age. MEASUREMENTS AND RESULTS: Hypercapnia increased the time spent in wakefulness and hypoxia the time spent in quiet sleep (QS). The state of alertness was the major determinant of HRV characterized with linear or nonlinear methods. Compared with QS, active sleep (AS) was associated with an overall increase in HRV magnitude and short-term self-similarity and a decrease in entropy of cardiac cycle length in air. This AS-related HRV pattern persisted in hypercapnia and was even more pronounced in hypoxia. CONCLUSION: Enhancement of AS-related sympathovagal coactivation in hypoxia, together with increased heart rate regularity, may be evidence that AS + hypoxia represent a particularly vulnerable state in early life. This should be kept in mind when deciding the optimal arterial oxygenation target in newborns and when investigating the potential involvement of hypoxia in SIDS pathogenesis.

Validation of a new automatic smoking machine to study the effects of cigarette smoke in newborn lambs.

The aim of this study was to describe the characteristics and validate the use of a new, custom-built automatic smoking machine (ASM), primarily designed to study the effects of an environmental tobacco smoke surrogate (ETS surrogate) exposure in animals of various sizes, including large animals. The equipment includes a programmable ASM coupled to a vented whole body chamber, where animals can be exposed to both mainstream and sidestream smoke. The user-friendly interface allows for full programming of puff volume (1-60 mL), time interval between two puffs (1-60 s) and between two cigarettes (1-60 min). Eight newborn lambs were exposed to either 10 (4 lambs, C10 group) or 20 (4 lambs, C20 group) cigarettes, 8 h per day for 15 days. Four additional control, lambs were exposed to air (C0 group). Weight gain was identical in all three groups of lambs. Urinary cotinine/creatinine ratio increased with the number of cigarettes smoked (C0: 11 ± 7 ng/mg; C10: 961 ± 539 ng/mg; C20: 1821 ± 312 ng/mg), with levels in the C10 and C20 groups in keeping with values published in infants exposed to ETS. Overall, results show that our new ASM is especially well suited for ETS surrogate exposure in non-restrained, non-anaesthetized large animals such as sheep.

Effects of postnatal smoke exposure on laryngeal chemoreflexes in newborn lambs.

Laryngeal chemoreflexes (LCR), which are elicited by the contact of liquids such as gastric refluxate with laryngeal mucosa, may trigger some cases of sudden infant death syndrome. Indeed, while LCR in mature mammals consist of protective responses, previous animal data have shown that LCR in immature newborns can include laryngospasm, apnea, bradycardia, and desaturation. The present study was aimed at testing the hypothesis that postnatal exposure to cigarette smoke is responsible for enhancing cardiorespiratory inhibition observed with LCR. Eight lambs were exposed to cigarette smoke (20 cigarettes/day) over 16 days and compared with seven control lambs. Urinary cotinine/creatinine ratio was measured at a level relevant to previously published levels in infants. On days 15 and 16, 0.5 ml of HCl (pH 2), milk, distilled water, or saline was injected onto the larynx via a chronic supraglottal catheter during sleep. Results showed that exposure to cigarette smoke enhanced respiratory inhibition (P < 0.05) and tended to enhance cardiac inhibition and decrease swallowing and arousal during LCR (P < 0.1). Overall, these results were observed independently of the state of alertness and the experimental solution tested. In conclusion, 16-day postnatal exposure to cigarette smoke increases cardiorespiratory inhibition and decreases protective mechanisms during LCR in nonsedated full-term lambs.

Postnatal autonomic activity in the preterm lamb.

BACKGROUND: Early postnatal abnormalities of autonomic nervous system (ANS) activity, including at baseline and following cardiorespiratory challenge, are involved in apneas-bradycardias of prematurity, apparent life-threatening events of infancy and sudden infant death syndrome. Literature data suggest that baseline ANS activity does not mature normally after premature birth. OBJECTIVES: This study performed in preterm lambs was aimed at assessing ANS maturation at baseline and following laryngeal chemoreflexes (LCR), a group of reflexes triggered by the contact of liquids with the laryngeal mucosa. METHODS: Heart rate variability (HRV) and baroreflex sensitivity (BRs) were measured at baseline and after LCR during polysomnographic recordings performed in five non-sedated lambs born 15 days prematurely. Laryngeal chemoreflexes were induced by distilled water or acid (pH 2) during sleep and wakefulness on postnatal days 7 (D7) and 14 (D14, full-term equivalence). RESULTS: While the life-threatening cardiorespiratory events of the LCR observed at D7 were no longer present at D14, baseline and post-LCR HRV and BRs indices were significantly lower at D14 compared to D7 (up to p<0.001). These results suggest that an initial autonomic overactivity was present at D7 and normalized at D14. CONCLUSION: The autonomic cardiac and baroreflex control appears to follow a specific evolution in the preterm compared to the full-term newborn lamb, with an important initial autonomic stress, which normalizes at an age equivalent to full-term. Potential relevance of these findings in relation to anomalies of cardiac control in the early postnatal period, such as apneas-bradycardias of prematurity, apparent life-threatening events of infancy and sudden infant death syndrome, awaits further studies.