Veratrine-induced reflexes and cough.

With cats anaesthetized with sodium pentobarbital we studied how veratrine-induced reflexes interact with cough. Cough was elicited by mechanical stimulation of tracheobronchial mucosa and its intensity was evaluated from the changes in oesophageal pressure. Veratrine injected intravenously produced apnoea, bradycardia and long-lasting hypotension. With each dose the intensity of cough was significantly decreased during the apnoea. When the mechanical stimulus was repeated during the breathing following apnoea with remaining hypotension, cough intensity parameters were not changed from control. Veratrine injected intracardially caused bradycardia, hypotension, and decreases in respiratory rate and tidal volume. The intensity of cough elicited just after injection of veratrine was also significantly decreased. We suggest that veratrine-induced reflexes depress the cough reflex mainly by inhibitory reflexes arising from cardiac receptors. The inhibition of cough is probably mediated indirectly via the inhibition of medullary respiratory neurons.

Components responsible for the surface tension of human tears.

PURPOSE: It was previously thought that the surface tension of tears was due to dissolved mucin, but it has recently been shown that very little mucin is present. The surface tensions of solutions of commercial mucin, lysozyme, lactoferrin or secretory IgA are all higher than that of tears. The influence of tear lipocalin and lipids remained to be tested. METHODS: Surface tension was determined by a micro-method on pooled intact stimulated human tears, and following extraction with lipid solvents. The extracted material was also added back, as was a variety of lipid standards (phospholipids, glycolipids, sterols, etc.). TLC and GLC were used in partial identification of the extract. Another lipocalin, bovine beta-lactoglobulin, was also tested alone and mixed with tear lipids, model lipids, or model tear proteins. RESULTS: Intact tears had a surface tension of 42-46 mN/m, but after lipid extraction this rose to 53-55.5 mN/m. Addition of lipids to the delipidised tear fluid gave a range of tensions from 42 to 49 mN/m, with the greatest effects shown by phospholipids (phosphatidylcholine, sphingomyelin), but full recovery was only achieved by using the extracted lipid material. Human meibomian oil was less effective. The GLC peak profile of the extract was markedly different from meibomian oil, and the TLC pattern was consistent with the presence of glycolipids. CONCLUSIONS: The surface tension of tears is due to a complex of tear lipocalin with a polar lipid fraction extractable from tears by lipid solvents and different from meibomian lipid. Lipocalin and this lipid fraction may be secreted together by the lacrimal gland.

[Functional and morphological changes in the respiratory organs of cats after long-term exposure to pure normobaric oxygen]. / Funkcné a morfologické zmeny dýchacích orgánov maciek dlhodobo exponovaných cistému normobarickému kyslíku.

BACKGROUND: Pulmonary oxygen toxicity is very well known and proved. The influence of hyperoxia on the respiratory reflexes is not known till now. MAIN PURPOSE: To ascertain if long-lasting breathing of pure normobaric oxygen (PNO) alter respiratory reflexes. METHODS: 34 adult cats of both sexes, weighing 2.5-4.0 kg, were used in two experiments. In the first experiment 16 animals with inserted chronic tracheal cannula (CTC) were used. In the second experiment 18 animals without CTC were employed. Part of animals in both experiments was exposed to PNO (day by day for 2 weeks, 10 h daily), remaining animals were exposed to room air under the same conditions. Side tracheal pressure was recorded in unanesthetized animals of the first experiment. The second experiment was performed in anaesthetised animals (Pentobarbital Spofa, 35 mg/kg, i.p.). Oesophageal pressure and blood pressure in femoral artery were recorded. Cough reflex, sneezing and aspiration reflexes were induced by mechanical stimulation of airway mucosa. Pulmonary chemoreflex was elicited by i.v. administration of 50 micrograms phenyl biguanid. Hering-Breuer inflation reflex was induced by lung inflation with pressure of 1 kPa. Reactivity of tracheal and pulmonary smooth muscle to histamine were measured in vitro. Differences in recorded parameters between animals exposed to PNO, and to room air, were tested by Mann-Whitney-Wilcoxon test and by Student's t-test. When p < 0.05, the differences were recognized as significant. RESULTS: Significant decreasing of the expiratory parameters of the cough induced from laryngopharyngeal mucosa, inhibition of sneezing, and inhibition of aspiration reflex, were found in animals exposed to PNO. Relaxing reaction of tracheal smooth muscle of control animals to histamine was reversed to contraction in animals exposed to PNO. Morphological changes of the respiratory tract induced by influence of oxygen were found CONCLUSION: Long-lasting breathing of PNO induced changes of respiratory reactions elicited mainly from upper airway.(Fig. 6, Tab. 3, Ref. 22)

Effects of midazolam and flumazenil on ventilation during sustained hypoxia in humans.

The purpose of this study was to investigate whether increases in gamma-aminobutyric acid (GABA) in the brain stem underlie the ventilatory decline observed during hypoxia in man. The ventilatory responses to sustained isocapnic hypoxia were studied in six adult male subjects on three separate days in three pharmacological conditions: (1) without any drug administration; (2) during infusion of midazolam (a drug which potentiates the effect of GABA); and (3) during infusion of flumazenil (a benzodiazepine antagonist). On each experimental day, the following protocol was repeated three times: end-tidal PO2 was held at 100 Torr for 10 min, then at 50 Torr for 20 min and finally at 100 Torr for 5 min. End-tidal PCO2 was held constant throughout. Responses in the three pharmacological conditions were similar. We conclude that neither potentiation of GABA transmission (midazolam) nor antagonism of this potentiation (flumazenil) greatly affect the decline in ventilation which occurs during extended exposure to hypoxia.

Effect of low-dose enflurane on the ventilatory response to hypoxia in humans.

To investigate the effects of enflurane on the control of breathing we have studied the ventilatory responses to isocapnic hypoxia in 12 adults with and without sedation with enflurane. Design 1 consisted of three steps into hypoxia (PE' O2 = 6.7 kPa), each lasting 3 min, separated by periods of euoxia lasting 5 min (PE' O2 = 13.3 kPa). Design 1 was repeated four times in each subject on the same day in random order: with carrier gas (control) and with 0.04 MAC, 0.07 MAC and 0.13 MAC of end-tidal enflurane concentrations. Design 2 consisted of 20-min exposures to hypoxia with and without 0.07 MAC of enflurane. Each exposure was preceded and followed by 5 min of euoxia. End-tidal PCO2 was held constant at 0.13-0.27 kPa greater than the resting level throughout both designs. Mean (SEM) ventilatory responses to hypoxia for design 1 were: 8.2 (1.3) litre min-1 (control), 6.6 (1.4) litre min-1 (0.04 MAC), 5.7 (1.1) litre min-1 (0.07 MAC) and 3.7 (0.5) litre min-1 (0.13 MAC) (P < 0.001). For design 2, enflurane produced a 15% reduction in resting ventilation (P < 0.01), a 40% decrease in the acute ventilatory response to hypoxia (P < 0.01) and a 32% reduction in ventilatory decline (ns) which occurred during sustained hypoxia.

Inhibition of active sodium absorption leads to a net liquid secretion into in vivo rabbit lung at two levels of alveolar hypoxia.

Active sodium transport across alveolar epithelium is known to contribute to the resolution of pulmonary oedema. We have attempted to assess whether sodium transport is essential to prevent liquid accumulation in healthy pulmonary alveoli exposed to mild hypoxia, and whether its contribution to liquid absorption differs between mild and moderate levels of hypoxia. In twenty-four anaesthetized adult rabbits we used direct bronchial cannulation to measure liquid movement from the liquid-filled left lung over 3.5 h. Half of the rabbits were studied at a level of mixed venous (and alveolar) oxygen partial pressure, PVO2, of 6.5 kPa and half at 4.5 kPa. PVO2 was altered by changing the inspired oxygen fraction in the ventilated right lung. Alveolar hydrostatic pressure was 0.3 kPa. In each group of 12, six animals with inhibitors of sodium transport in the isosmotic instillate were compared with six controls. We have shown an alveolar liquid secretion (approximately 0.6 microl min(-1) (kg body weight)(-1)) in the presence of inhibitors of active transport and an absorption (approximately 4 microl min(-1) (kg body weight)(-1)) in controls. Changing PVO2 had no influence on these movements. We conclude that, in this model of pulmonary oedema, active sodium transport appears to be essential for prevention of alveolar liquid accumulation via secretion. Furthermore, the contribution of active sodium transport to liquid absorption remains constant at oxygen tensions between 4.5 and 6.5 kPa.

Comparison of the effects of sub-hypnotic concentrations of propofol and halothane on the acute ventilatory response to hypoxia.

To compare the effects of sub-anaesthetic concentrations of propofol and halothane on the respiratory control system, we have studied the acute ventilatory response to isocapnic hypoxia (AHVR) in 12 adults with and without three different concentrations of propofol and halothane. Target doses for propofol were 0, 0.05, 0.1 and 0.2 of the effective plasma concentration (EC50 = 8.1 micrograms ml-1). Target doses for halothane were 0, 0.05, 0.1 and 0.2 minimum alveolar concentration (MAC = 0.77%). The doses achieved experimentally were 0.01, 0.06, 0.13 and 0.26 of the EC50 for propofol and 0, 0.05, 0.11 and 0.20 MAC for halothane. During the experiment subjects breathed via a mouthpiece from an end-tidal forcing system. End-tidal PO2 (PE'O2) was held at 13.3 kPa for 5 min, and then at 6.7 kPa for 5 min. End-tidal PCO2 (PE'CO2) was held constant at 0.13-0.27 kPa greater than the subject's natural level throughout. The mean values for AHVR with propofol were: 12.8 (SEM 2.4) litre min-1 (0.01 EC50), 10.0 (1.9) litre min-1 (0.06 EC50), 9.8 (2.3) litre min-1 (0.13 EC50) and 4.9 (1.2) litre min-1 (0.26 EC50). The values for AHVR with halothane were: 11.9 (2.4) litre min-1 (0 MAC), 7.8 (1.6) litre min-1 (0.05 MAC), 5.9 (1.2) litre min-1 (0.11 MAC) and 3.2 (1.6) litre min-1 (0.2 MAC). The decline in AHVR with increasing dose for both drugs was statistically significant (ANOVA, P < 0.001); there was no significant difference between the two drugs with respect to this decline. Normoxic ventilation with propofol declined from 13.2 (1.6) litre min-1 (0.01 EC50) to 8.3 (0.9 litre min-1 (0.26 EC50), and with halothane declined from 13.5 (2.0) litre min-1 (0 MAC) to 11.8 (1.6) litre min-1 (0.2 MAC). This was significant for both drugs (ANOVA, P < 0.001).

The human ventilatory response to step changes in end-tidal PO2 of differing amplitude.

This study assessed whether the form of the peripheral chemoreflex response to hypoxia depends on the magnitude of the stimulus. Two amplitudes of square-wave hypoxic stimulation were employed: small amplitude (SO) PETO2 from 63.2 to 54.9 Torr, and large amplitude (LO) PETO2 from 73.0 to 48.0 Torr. Each was studied at two levels of PETCO2: 2 Torr above resting PETCO2 (EC), and 7 Torr above resting PETCO2 (HC). Each protocol was repeated 6 times on 5 subjects. To assess the form of the response, a simple first-order model was fitted to the data which incorporated a pure delay (Td) and time constant (tau). Average parameter values (sec) were: ECSO tau = 4.07, Td = 6.69; ECLO tau = 8.82, Td = 4.91; HCSO tau = 5.22, Td = 7.08; HCLO tau = 9.96, Td = 4.39. ANOVA demonstrated modest but significant differences for loge(tau) (P < 0.01) and Td (P < 0.02) between the two hypoxic step magnitudes, with tau longer and Td shorter for the larger step size, but no differences were found between the parameter values for the two CO2 levels. We conclude that the form of the response of the peripheral chemoreflex to hypoxia depends upon the magnitude of the stimulus.

Dependence of pulmonary venous admixture on inspired oxygen fraction and time during regional hypoxia in the rabbit.

In order to examine the value of assuming constant pulmonary venous admixture with respect to changes in inspired oxygen fraction (FIO2) and time during sustained unilateral hypoxia, we studied venous admixture for 6 h in 27 anaesthetized rabbits in which the left lung was filled with liquid, isosmotic with plasma. In one group of 10 rabbits the right lung was ventilated for 6 h with FIO2 = 1; in a second group of 10 the right lung was ventilated with FIO2 = 1 for 2.5 h and then with FIO2 = 0.3 for 3.5 h. A third group was similarly studied by changing from FIO2 = 1 to FIO2 = 0.5. We found that hypoxic pulmonary vasoconstriction continued to intensify over 3 h. At 3-6 h, with FIO2 = 0.3, venous admixture (0.32 (SEM 0.03)) was higher than baseline (0.13 (0.01), t = 0 min during bilateral oxygenation) by twice the elevation above baseline of the venous admixture (0.22 (0.01)) in the group with FIO2 = 1. The finding of a marked increase in venous admixture with decreasing FIO2 is discussed in relation to current models of hypoxic pulmonary vasoconstriction.

Respiration and airway reflexes after transversal brain stem lesions in cats.

The effect of brain stem transection at different levels of the pons Varolii and the medulla oblongata on respiration and on cough and the aspiration and expiration reflex elicited by mechanical stimulation of the relevant parts of the respiratory tract was studied in experiments on 13 anaesthetized, unparalyzed cats. The results of 142 respiratory reflex elicitation tests showed that: 1. Compared with the control state, transection of the upper and middle part of the pons Varolii and transection at the level of the pontomedullary junction reduced the respiration rate (p less than 0.001), increased the duration of inspiration and expiration (p less than 0.001, transection 10 mm rostrally to the obex) and gave rise to apneustic breathing (8 mm), or to tonic, respiration-modulated activity of the phrenic nerve and diaphragm (6 mm). 2. Successive transection of the pons and the pontomedullary junction region led chiefly to a drop in maximum expiratory pleural pressure values (p less than 0.01-0.001) during cough and the expiration reflex and to a drop in maximum inspiratory pleural pressure values during the aspiration reflex (p less than 0.02-0.001). 3. Transection of the upper part of the medulla oblongata always led to permanent arrest of rhythmic respiration, during which cough and the expiration reflex could not be elicited while the aspiration reflex persisted (though in a weakened form). This state was followed by gasping, during which only a highly elicitable aspiration reflex persisted. 4. It can be assumed from the above findings that the central mechanisms responsible for the development of powerful expiratory efforts in cough and the expiration reflex could be localized in the pons Varolii, while those integrating the aspiration reflex are probably localized mainly in the medulla oblongata.

Influence of 0.2 minimum alveolar concentration of enflurane on the ventilatory response to sustained hypoxia in humans.

To determine the influence of 0.2 minimum alveolar concentration (MAC) of enflurane on the time course of ventilation during sustained hypoxia, we studied 10 healthy adult volunteers with and without enflurane. The following design was used: end-tidal Po2 was maintained at 13.3 kPa for 8 min, at 6.7 kPa for 20 min and at 13.3 kPa for 8 min. End-tidal Pco2 was held constant throughout at 0.67 kPa above the subject's natural value. Control experiments were conducted with no hypoxia imposed. During the experiment subjects breathed via a mouthpiece from an automated gas mixing system which controlled end-tidal values. Enflurane reduced baseline (euoxic) ventilation from 20.9 (SEM 2.0) litre min-1 to 10.1 (1.0) litre min-1 (ANOVA, P < 0.001). Enflurane reduced the acute ventilatory response to hypoxia (AHVR) from 20.1 (3.3) litre min-1 to 5.0 (1.3) litre min-1 (ANOVA, P < 0.01), and the ventilatory off-response at cessation of hypoxia from 11.7 (2.4) litre min-1 to 1.8 (0.5) litre min-1 (ANOVA, P < 0.02). There was no significant difference in hypoxic ventilatory decline (HVD) without and with enflurane (8.9 (2.4) litre min-1 vs 5.5 (1.1) litre min-1; ANOVA, ns). These results confirm that 0.2 MAC of enflurane suppressed the acute ventilatory response to hypoxia, but had no significant effect on the subsequent ventilatory decline during sustained hypoxia.

Time course of hypoxic pulmonary vasoconstriction: a rabbit model of regional hypoxia.

There is disagreement in the literature about the time required for hypoxic constriction of pulmonary vessels to reach its full intensity. Some studies suggest that only minutes are required, others that several hours are needed. We examined the time course over 6 h of changes in pulmonary shunt (as a fraction of cardiac output) following induction of unilateral hypoxia by collapse or liquid filling of the left lung in 47 anesthetized rabbits. The time course was examined at four degrees of lung inflation: during collapse and at airway pressures of 0.3 kPa, 0.6 kPa, and 0.9 kPa. The respective volumes (mean +/- SD) of the liquid-filled lung were estimated to be 6.4 +/- 1.0, 12.8 +/- 2.5, and 15.8 +/- 1.6 ml/kg body weight (BW). During sustained hypoxia (the period from 150 to 360 min after inducing hypoxia), shunt declined at a slow linear rate of 2.37 x 10(-4)/min, which was independent of lung inflation (p = 0.65 analysis of variance [ANOVA]) and significantly different from zero (p < 0.001). The stability of cardiac output in this animal model, as measured sequentially by thermodilution, was confirmed in a further 20 animals. The experiments provide evidence for a slow intensification of blood-flow diversion at a rate that does not depend upon the degree of lung inflation. Whether this change is a feature of hypoxic constriction itself, or some modulation of it, remains unclear.

Physical properties of stimulated and unstimulated tears.

It has long been assumed that unstimulated tears are more thoroughly equilibrated with epithelial secretions than stimulated tears, since they are in contact with tarsal, bulbar and corneal surfaces for longer. It was also believed from results with model solutions that soluble mucin is responsible for the observed surface tension and viscosity of tears. If longer contact means more mucin is dissolved in the aqueous tears, then the surface activity (surface tension lowered by mucin) and viscosity (raised by mucin) of tears should therefore be enhanced in unstimulated over stimulated tears. Pools of stimulated and minimally-stimulated tears were collected from a group of healthy adult volunteers by glass capillary. Viscosities were measured in the Contraves Low Shear 30 rheometer over the range of shear rates 0-130 sec-1. Surface tension was measured in the collection capillaries by a micro-technique, before and after refrigerated storage. Both surface tension and viscosity were determined for a variety of tear proteins and mucins. No significant difference was found between the viscosity/shear rate plots of stimulated and unstimulated tear samples. The viscosities of solutions of individual tear proteins were low, except for the combination of lysozyme and secretory IgA. Surface tensions were also similar in both cases, and unchanged by storage at room temperature or refrigeration, indicating no significant loss of surface-active material by adsorption on the capillary walls. Results with model mucin solutions gave a variety of results indicating either little surface activity or losses due to wall adsorption. Tear proteins, individually or in combination, did not lower surface tension to the level of tears. Tear viscosity seems not to depend on the level of dissolved mucins. This suggests either that a constant level of these is picked up even by short-term contact with ocular surfaces, or that viscosity arises from currently unknown materials which vary little with tear flow rate. This type of shear-dependent viscosity is most easily simulated in model solutions with polyionic linear macromolecules, including mucins. The contribution of individual proteins to overall viscosity is small, but combinations including lysozyme show tear-like characteristics, and may indicate that proteins whose concentration is relatively independent of tear flow rate combine with other tear components (possibly including mucins or lipids) to produce their full effect on tear viscosity. The surface tension results suggest that mucins are not of primary importance. Theories of tear film structure and performance need revision.

Effects of focal cooling of medulla oblongata structures on quiet breathing in cats.

Experiments were carried out on 16 anaesthetized, non-paralysed cats to determine the effects of unilateral, successive focal cooling of the nuclei of the dorsal and ventral respiratory groups (DRG, VRG) of the medulla oblongata on quiet breathing parameters. The results of cold block tests of the respiratory nuclei showed that: 1. Compared with the control state, cooling of the ventrolateral part of the nucleus solitarii (vl. NTS) and the rostral part of the nucleus retroambigualis (r. NRA) to 20 degrees C or 15 degrees C decreased the respiration rate (p less than 0.001), prolonged the inspiration time (p less than 0.01 and p less than 0.001 respectively) and the development of apneustic breathing. A decrease in the inspiratory pleural pressure values (p less than 0.01) was found after cooling the r. NRA region to 15 degrees C. In 45% of the cases of cooling of the vl. NTS and 66.7% of cooling of the r. NRA to 15 degrees C, an incidence of short inspiratory efforts was observed. 2. Focal cooling of the nucleus retrofacialis (nucl. RF) region to 20 degrees C always arrested rhythmic respiration. 3. The effects of unilateral focal cooling of the respiratory nuclei were always bilaterally symmetrical and, after discontinuing cooling, reversible. 4. The findings indicate that the inspiratory neurones of the r. NRA participate more in regulation of the intensity of inspiration than those of the vl. NTS, while the nucl. RF region may be a part of central regulatory mechanisms essential for the maintenance of rhythmic breathing in cats.