On the pathway of an animal model for restless legs syndrome...

Restless legs syndrome (RLS) is a chronic sleep motor disorder that affects up to 10% of the general population. Except for periodic leg movements (PLM), which can be found in the great majority of RLS patients, no objective hematochimic or neurophysiological markers are available to prove the diagnosis, which is based on clinical standard criteria. Nowadays, the aetiopathogenesis of the syndrome is unknown. In a consistent sample of patients affected by the idiopathic form, the disease is inherited as an autosomal dominant trait related to an unidentified locus, while each symptomatic form is probably linked to a specific cause. Although of possible different origins, both the primary and secondary forms may share the same pathogenetic mechanism, which, even if unclear, could be characterised by a neurological dysfunction of the dopaminergic system. Several issues, including strong efficacy of dopamine-agonist treatments, support this theory, which is currently considered the main pathogenetic hypothesis. Most of the past studies tried to clarify the RLS mechanism using the neurophysiological, biochemical and neuroimaging techniques applied to the field of human research. Now the time has come to accept the challenge in creating an animal model of RLS, which may emerge as a decisive step in understanding RLS pathogenesis, and to develop and test new therapies. Even though there have been a few significant efforts, a valid animal model of RLS still does not exist. In past pioneering studies, the authors attempted to induce restless motor behaviour in animals by different strategies: antidopaminergic pharmacological interventions, spinal or cerebral lesions of specific regions involved in the motor control and in dopamine regulation, and selective deletion of genes coding for dopamine receptors. Rodents (mice and rats) were always chosen by the authors as the animals for their experiments. The current tendency in achieving an RLS model is generally represented by simulation of a symptomatic condition of RLS or by a direct interference of the dopaminergic system. In this regard, the pharmacological method had the intention to reproduce the neuroleptic-induced acathisia, the spinal lesional model was based on the hypothesis of myelopathic- related PLM, and the hypothalamic lesion tested the motor consequence of A11 dopaminergic neurons. Preliminary studies are underway to replicate the pregnancy-related form of RLS by using a hormonal intervention, and the iron-deficiency secondary form by using specific iron-free diets. Today, modern technologies are available to easily replicate in animals most of the symptomatic RLS conditions. In addition, more than a few well validated animal models of different diseases known to be related to RLS or PLM, for instance, Parkinson's disease, rheumatoid arthritis and renal failure, could also be exploited in addressing this topic. The real obstacle in achieving an RLS model is the absence of a certain diagnostic marker to recognise if the animal that underwent the different experimental procedures has developed the RLS condition or not. Concerning this issue, possible specific endpoints are represented by the increase in locomotor activity, which are ascertainable by different techniques, such as openfield or run-wheel activity, or by sleep fragmentation, in which the circadian shift can be verified by applying polysomnography on the animal. PLM are probably the only specific and reliable markers available to recognise and quantify experimentally induced RLS. Despite a few authors who reported the presence of limb-phasic, pseudoperiodic activity during sleep in old or in lesioned rats, the existence of spontaneous or provoked PLM in animals is still debated. Eventually, the PLM features in an animal could be markedly different compared to human ones. To recognise and characterise PLM in animals, three more essential steps are required: a method to record directly, as in humans, the activity of the tibialis anterior (TA) muscles, a consistent amount of normative control data on the TA activity in healthy animals, and reliable analysis to distinguish the generic phasic muscular activity to a possible unambiguous PLM pattern. This review includes a summary and a critical discussion of the previous tentative RLS models, proposals for other possible animal models, and firstly the preliminary normative data on TA activity during sleep in normal rodents.

Reciprocal activation of hypopharyngeal muscles and their effect on upper airway area.

We examined in awake goats, 1) with intact upper airways (UAW), the effect of altering chemical drive on pharyngeal constrictors [thyropharyngeus (TP) and hypopharyngeus (HP)] and a dilator [stylopharyngeus (SP)], and 2) with an isolated UAW, the effect of activation of these muscles on supraglottic UAW (UAW(SG)) area. During eupnea in nine goats with intact UAW, the TP and HP were active during expiration, whereas the SP exhibited tonic expiratory and phasic inspiratory activity. After mechanically induced apneas (MIA), TP activity increased (263%, P < 0.02), HP activity exhibited a small, varied response, and SP activity greatly decreased (10%, P < 0.02). During resumption of respiratory effort, all goats exhibited absent/reduced airflow, and when diaphragm activity was 95% of control, TP activity remained elevated (135%) and SP activity was reduced (56%, P < 0.02). During hypercapnia, 1) TP activity decreased (P < 0.02), 2) HP response varied, and 3) SP activity increased (P < 0.02). After MIA in six goats with isolated UAW, TP activity increased 198% (P < 0.02) and UAW(SG) area (endoscopically determined) decreased (to 15% of control, P < 0.02). During recovery from MIA, a correlation was found between UAW(SG) area and the ratio of SP to TP activity. We conclude that the reciprocal activation of mechanically opposing dilator and constrictor muscles in the hypopharynx is correlated to changes in the UAW(SG) area, and an imbalance in activity of these opposing muscles can lead to UAW(SG) narrowing.

Interaction of cross-sectional area, driving pressure, and airflow of passive velopharynx.

Previous studies have shown that, when the pharyngeal muscles are relaxed, the velopharynx is a highly compliant segment of the pharynx. Thus, under these circumstances, cross-sectional area of the velopharynx (AVP), driving pressure across the velopharynx (DeltaP), and inspiratory airflow (VI) will be mutually interdependent variables. The purpose of the present investigation was to describe the interrelation among these three variables during inspiration. We studied 15 sleeping patients with obstructive sleep apnea/hypopnea when the pharyngeal muscles were rendered hypotonic by applying continuous positive airway pressure to the nasal airway. AVP, determined by endoscopic imaging, was significantly greater at onset of VI limitation than at minimum oropharyngeal pressure (P < 0. 01). Snoring was never observed during VI limitation. In a subgroup of six patients, values for DeltaP, VI, and AVP were obtained at 0. 1-s intervals at various levels of mask pressure. For these six patients, the mathematical expression VI = 0.657(AVP/Amax) . DeltaP0. 332, where Amax is maximal AVP, described the relationship among the three variables (R2 = 0.962) for flow-limited and non-flow-limited inspirations. The impedance of the passive velopharynx, defined as DeltaP0.33/V, was inversely related to AVP and increased dramatically when AVP was <0.3 cm2. In summary, we observed a progressive decrease in AVP during flow-limited inspiration in patients with obstructive sleep apnea. This constriction of the velopharynx contributes to an increase in velopharyngeal impedance that, in turn, counterbalances the increase in DeltaP during flow limitation.

Negative pressure effects on mechanically opposing pharyngeal muscles in awake and sleeping goats.

Our aim was to investigate the effects of the negative pressure reflex on mechanically opposing pharyngeal muscles during wakefulness, slow-wave sleep (SWS), and rapid eye movement (REM) sleep. In four goats with isolated upper airways, we measured tracheal airflow and electrical activity of the thyropharyngeus (TP; constricting), the stylopharyngeus (SP; dilating), and the diaphragm (Dia). In the wakefulness state in response to negative pressure tests, TP decreased (65%), SP increased (198%), and tidal volume (VT) (66%) and rate of rise of Dia (Dia(slope), 69%) decreased (P < 0.02). Similarly, during SWS, the negative pressure response of TP (31%), VT (61%), and Dia(slope) (60%) decreased, whereas SP (113%) increased, relative to SWS control (P < 0.02). In REM sleep, the negative pressure response by TP and SP were small, whereas both VT (38%) and Dia(slope) (24%) were greatly decreased (P < 0.02) compared with REM control. Inspiratory duration remained unchanged in response to negative pressure tests in all states. These data provide evidence that mechanically opposing inspiratory and expiratory pharyngeal muscles are reciprocally controlled and their response to negative pressure are state dependent.

Static mechanics of the velopharynx of patients with obstructive sleep apnea.

The static mechanics of the hypotonic pharynx were endoscopically evaluated in nine sleeping patients with obstructive sleep apnea, having a primary narrowing only at the velopharynx. The velopharynx closed completely at a mean pressure of 0.18 +/- 1.21 cmH2O, and the mean half-dilation pressure was 1.93 cmH2O above closing pressure. The dependence of area on pressure was distinctly curvilinear, being steep near closing pressure and asymptotically approaching maximum area (mean = 1.32 cm2). The data for each patient were satisfactorily fitted by an exponential function (mean R2 = 0.98), and a single exponential relationship usefully represented the dependence of relative area on pressure above closing pressure for the population (R2 = 0.85). During the test inspiration, flow limitation was consistently observed when mask pressure exceeded closing pressure by 0.5-3.0 cmH2O. In summary, the static mechanics of the hypotonic velopharynx of patients with obstructive sleep apnea can be described by an exponential pressure-area relationship, with a closing pressure near atmospheric pressure and a high compliance in the range of airway pressure 0-3 cmH2O above closing pressure.

Multiple rostral medullary nuclei can influence breathing in awake goats.

The purpose of this study was to determine the effect on breathing of neuronal dysfunction in the retrotrapezoid (RTN), facial (FN), gigantocellularis reticularis (RGN), or vestibular (VN) nuclei of adult awake goats. Microtubules were chronically implanted to induce neuronal dysfunction by microinjection of an excitatory amino acid (EAA) receptor antagonist or a neurotoxin. The EAA receptor antagonist had minimal effect on eupneic breathing, but 8--10 days after injection of the neurotoxin, 7 of 10 goats hypoventilated (arterial PCO(2) increased 3.2 +/- 0.7 Torr). Overall there were no significant (P > 0.10) effects of the EAA receptor antagonist on CO(2) sensitivity. However, for all nuclei, > or =66% of the antagonist injections altered CO(2) sensitivity by more than the normal 12.7 +/- 1.6% day-to-day variation. These changes were not uniform, inasmuch as the antagonist increased (RTN, n = 2; FN, n = 7; RGN, n = 6; VN, n = 1) or decreased (RTN, n = 2; RGN, n = 3; VN, n = 2) CO(2) sensitivity. Ten days after injection of the neurotoxin into the FN (n = 3) or RGN (n = 5), CO(2) sensitivity was also reduced. Neuronal dysfunction also did not have a uniform effect on the exercise arterial PCO(2) response, and there was no correlation between effects on CO(2) sensitivity and the exercise hyperpnea. We conclude that there is a heterogeneous population of neurons in these rostral medullary nuclei (or adjacent tissue) that can affect breathing in the awake state, possibly through chemoreception or chemoreceptor-related mechanisms.

Large lesions in the pre-Bötzinger complex area eliminate eupneic respiratory rhythm in awake goats.

In awake goats, 29% bilateral destruction of neurokinin-1 receptor-expressing neurons in the pre-Bötzinger complex (pre-BötzC) area with saporin conjugated to substance P results in transient disruptions of the normal pattern of eupneic respiratory muscle activation (Wenninger JM, Pan LG, Klum L, Leekley T, Bastastic J, Hodges MR, Feroah T, Davis S, and Forster HV. J Appl Physiol 97: 1620-1628, 2004). Therefore, the purpose of these studies was to determine whether large or total lesioning in the pre-BötzC area of goats would eliminate phasic diaphragm activity and the eupneic breathing pattern. In awake goats that already had 29% bilateral destruction of neurokinin-1 receptor-expressing neurons in the pre-BötzC area, bilateral ibotenic acid (10 microl, 50 mM) injection into the pre-BötzC area resulted in a tachypneic hyperpnea that reached a maximum (132 +/- 10.1 breaths/min) approximately 30-90 min after bilateral injection. Thereafter, breathing frequency declined, central apneas resulted in arterial hypoxemia (arterial Po2 approximately 40 Torr) and hypercapnia (arterial Pco2 approximately 60 Torr), and, 11 +/- 3 min after the peak tachypnea, respiratory failure was followed by cardiac arrest in three airway-intact goats. However, after the peak tachypnea in four tracheostomized goats, mechanical ventilation was initiated to maintain arterial blood gases at control levels, during which there was no phasic diaphragm or abdominal muscle activity. When briefly removed from the ventilator (approximately 90 s), these goats became hypoxemic and hypercapnic. During this time, minimal, passive inspiratory flow resulted from phasic abdominal muscle activity. We estimate that 70% of the neurons within the pre-BötzC area were lesioned in these goats. We conclude that, in the awake state, the pre-BötzC is critical for generating a diaphragm, eupneic respiratory rhythm, and that, in the absence of the pre-BötzC, spontaneous breathing reflects the activity of an expiratory rhythm generator.

Effects on breathing in awake and sleeping goats of focal acidosis in the medullary raphe.

Our aim was to determine the effects of focal acidification in the raphe obscurus (RO) and raphe pallidus (RP) on ventilation and other physiological variables in both the awake and sleep states in adult goats. Through chronically implanted microtubules, 1) a focal acidosis was created by microdialysis of mock cerebrospinal fluid (mCSF), equilibrated with various levels of CO2, and 2) medullary extracellular fluid (ECF) pH was measured by using a custom-made pH electrode. Focal acidosis in the RO or RP, by dialyzing either 25 or 80% CO2 (mCSF pH approximately 6.8 or 6.3), increased (P < 0.05) inspiratory flow by 8 and 12%, respectively, while the animals were awake during the day, but not at night while they were awake or in non-rapid eye movement sleep. While the animals were awake during the day, there were also increases in heart rate and blood pressure (P < 0.05) but no significant change in metabolic rate or arterial Pco2. Dialysis with mCSF equilibrated with 25 or 80% CO2 reduced ECF pH by the same amount (25%) or three times more (80%) than when inspired CO2 was increased to 7%. During CO2 inhalation, the reduction in ECF pH was only 50% of the reduction in arterial pH. Finally, dialysis in vivo only decreased ECF pH by 19.1% of the change during dialysis in an in vitro system. We conclude that 1) the physiological responses to focal acidosis in the RO and RP are consistent with the existence of chemoreceptors in these nuclei, and 2) local pH buffering mechanisms act to minimize changes in brain pH during systemic induced acidosis and microdialysis focal acidosis and that these mechanisms could be as or more important to pH regulation than the small changes in inspiratory flow during a focal acidosis.

Posterior cephalometric radiographic analysis in obstructive sleep apnea.

Our objective was to evaluate the relationship between posterior facial cephalometric measures and obstructive sleep apnea syndrome (OSAS). We used a consecutive sample of 60 patients with OSAS who underwent upright lateral cephalograms, uvulopalatopharyngoplasty (UPPP), and preoperative and postoperative polysomnography. Successful responders to UPPP were arbitrarily defined as having a respiratory disturbance index reduced to fewer than 20 events per hour. Standard cephalometric measurements were used. Posterior facial height measures were constructed, based on a plane perpendicular to the Frankfort horizontal placed at hyoidale. The total and lower airway lengths were shorter and posterior mandibular height was longer in UPPP responders compared to nonresponders (p < or = .05). There was no difference between the two groups by standard cephalometric measurements. Responders and nonresponders to UPPP have significant differences in posterior airway measures that are not reflected in standard cephalometric measures. Airway length likely is a critical factor in OSAS and surgical response.

Relationship of the head impulse test and head-shake nystagmus in reference to caloric testing.

OBJECTIVE: The objective of this study was to determine the usefulness of the head impulse test (HIT) and head-shake nystagmus (HSN), two easily performed office maneuvers, in the evaluation of the dizzy patient with reference to caloric irrigation results. OBJECTIVE AND SETTING: This was a prospective double-blind trial conducted at an outpatient academic tertiary referral center. PATIENTS: The study population was composed of 105 patients (35 male, 70 female) who presented for evaluation of dizziness and ranged in age from 13 to 87 years (mean 52.1). INTERVENTION: The intervention was HIT and HSN evaluation followed by bithermal binaural air caloric irrigations. MAIN OUTCOME MEASURES: The main outcome measures were sensitivity, specificity, and predictive values of HIT and HSN evaluation (individually and in combination) in relation to caloric results. RESULTS: Sensitivity of the tests was equally low (35%), whereas specificity was high (HIT 95%, HSN 92%). The positive predictive value for the two tests in combination (80%) was greater than for each individually (HIT 64%, HSN 50%). Negative predictive values remained stable when considering each test individually (HIT 86%, HSN 86%) or in combination (88%). CONCLUSIONS: The low sensitivity renders both tests inadequate as a screening tool for peripheral vestibular disease based on caloric results. However, when HIT and HSN results are both abnormal, there is a high likelihood of a significant caloric deficit.

Effect of slow wave and REM sleep on thyropharyngeus and stylopharyngeus activity during induced central apneas.

The pharyngeal constrictors have been hypothesized to play an important role in the regulation of upper airway (UAW) patency in patients with sleep apnea. However, little research has focused on the activation and control of muscles that determine the lateral and posterior wall of the retropalatal airway dimensions. Our aim was to investigate the effects of slow wave sleep (SWS) and rapid eye movement (REM) sleep on the activation of pharyngeal constrictor (thyropharyngeus; TP) and dilator (stylopharyngeus; SP) muscles during eupneic breathing and induced central apneas. In nine goats, we found that eupneic TP and SP activity progressively decreased from awake to SWS (57 and 56%, respectively; P<0.01) and further in REM (25.6 and 19.9%, respectively; P<0.01). In contrast, diaphragm activity decreased equally during SWS and REM (89.3 and 87.7%, respectively; P<0.01) compared to awake. Following induced apneas while SP activity was eliminated in every state, maximal TP activity was highest in awake state (318.6% of control; P<0.02), less in SWS (157.6%; P<0.02), and nearly absent in REM (117.3%; P>0.02). During the recovery from an induced apnea when diaphragm activity was at 95% of its' control, awake TP activity remained significantly elevated and SP reduced (P>0.02) while TP activity during SWS was elevated and SP had returned to control level. During REM, TP and SP activity were not different from their reduced controls (P>0.02). The data supports our hypotheses that SWS and REM sleep causes a reduction in the eupneic TP and SP activity, as well as a reduction in TP response to induced apneas. However, the relative imbalance in TP vs SP activity during the recovery from an apnea (awake and SWS) suggest that an imbalance of active neuromuscular forces may contribute to upper airway narrowing in mixed apneas, but not in central apnea during sleep.

Pharyngeal narrowing and closing pressures in patients with obstructive sleep apnea.

Based on previous studies, we hypothesized that the pharynx collapses at multiple sites in most patients with obstructive sleep apnea (OSA). The purpose of this study was to document, in a population of apneic subjects, the site(s) of narrowing and closing pressure of the hypotonic pharynx. We endoscopically examined the pharynx in 45 OSA patients during sleep while they received nasal continuous positive airway pressure (CPAP), which produces hypotonia of pharyngeal muscles. Intrapharyngeal images and pressures were obtained at the end of expiration during single-breath tests (SBT). The fractional narrowing (FN) of each pharyngeal segment (nasopharynx, oropharynx, and hypopharynx) was calculated as the relative change in area when nasal airway pressure was reduced from a pressure that held the pharynx fully distended to the pressure at which the airway closed. The frequency distribution of FN for the nasopharynx was skewed toward larger values, and the frequency was relatively evenly distributed for the oropharynx and hypopharynx. A site having FN greater than 0.75 was defined as a site of primary narrowing, and a site showing FN 0.25 to 0.75 was defined as a site of secondary narrowing. The nasopharynx was a site of primary narrowing in 80% of patients, and two or more sites of narrowing were commonly observed (82%). Four categories of combined narrowing were identified: (1) primary narrowing only at the nasopharynx (18%); (2) primary narrowing at the nasopharynx plus other sites of secondary narrowing (40%); (3) primary narrowing at the nasopharynx plus other sites of primary narrowing (22%); and (4) other patterns (20%).(ABSTRACT TRUNCATED AT 250 WORDS)

Site of pharyngeal narrowing predicts outcome of surgery for obstructive sleep apnea.

Uvulopalatopharyngoplasty (UPPP), an operation that enlarges the pharyngeal airway at the level of the soft palate, improves respiratory status during sleep in only 50% of patients with obstructive sleep apnea (OSA). This poor outcome suggests that narrowing of the pharyngeal airway at nonpalatal sites contributes to the obstructive process in many patients with OSA. We have used a novel endoscopic method to identify regions of the passive pharyngeal airway most susceptible to narrowing or complete closure. In order to test the hypothesis that narrowing of the passive airway at the nasopharynx predicts a favorable surgical outcome, we have preoperatively assessed the local mechanics of the passive pharyngeal airway in 18 patients with OSA undergoing UPPP. The patient population was prospectively divided into two groups: an exclusively nasopharyngeal (ENP) group, consisting of patients exhibiting narrowing only in the nasopharynx, and a not exclusively nasopharyngeal (NENP) group, consisting of patients having at least one site of narrowing outside the nasopharynx. The frequency of respiratory disturbances and arousals and the cumulative time in apnea-hypopnea were significantly reduced after surgery for the ENP group, but not for the NENP group. Improvement rate for the ENP group (86%) exceeded that for the NENP group (18%) (p < 0.01). These differences became even greater when selection criteria for the ENP group were made more restrictive (i.e., restricted to the velopharynx) or more liberal (i.e., including secondary narrowing of the oropharynx). Our results show that evaluation of passive pharyngeal mechanics identifies patients with OSA likely to improve after UPPP.